Compounds for activating invariant natural killer t-cells and methods of use in eliminating inflammatory senescent cells

ABSTRACT

Compounds for activating invariant natural killer T cells (iNKT) cells are provided. Compounds according to certain embodiments activate iNKT cells and induce an increase in the production of one or more cytokines, such as IFN-γ, IL-2, IL-4, IL-6 and TNFα. In some embodiments, activated iNKT cells are used to selectively reduce the presence of or eliminate inflammatory senescent cells, such as senescent cells having an inflammatory secretome (SASP). Methods for activating iNKT cells by contacting an iNKT cell with an amount of the subject compounds and selectively reducing the presence of or eliminating senescent cells with activated iNKT cells are also described. Compositions for practicing the subject methods are also described.

In a healthy system, the immune system naturally (endogenously) clearssenescent cells. When this immune function is compromised, senescentcells build up and can propagate into a multitude of different diseases.Invariant natural killer T (iNKT) cells are a subset of T cells thatrecognize glycolipid antigens bound to the cluster of differentiation(CD)1d molecule expressed by surface antigen presenting cells.Recognition of exogenous and endogenous lipids can aid in immuneresponse to maladies such as autoimmune disease, allergic disease,metabolic syndrome, cancer and pathogen infection. Although iNKT cellshave been shown to mediate immune responses based on cytokine release,iNKT cells can also function as effectors by cell cytotoxicity.

SUMMARY

Compounds for activating invariant natural killer T cells (iNKT) cellsare provided. Compounds according to certain embodiments activate iNKTcells and induce an increase in the production of one or more cytokines,such as IFNγ, IL-2, IL-4, IL-6 and TNFα. In some embodiments, activatediNKT cells are used to selectively reduce the presence of or eliminateinflammatory senescent cells, such as senescent cells having aninflammatory secretome (SASP). Methods for activating iNKT cells bycontacting an iNKT cell with an amount of the subject compounds andselectively reducing the presence of or eliminating senescent cells withactivated iNKT cells are also described. Compositions for practicing thesubject methods are also described.

In some embodiments, compounds of interest include a compound of formulaDCD-(I):

-   -   wherein:    -   Z is selected from:

-   -   wherein        indicates the Z—O bond;    -   wherein X is —NHCO— or oxygen;    -   R¹, R², R³ and R⁴ are each independently selected from hydrogen,        alkyl, substituted alkyl, heteroalkyl, substituted heteroalkyl,        cycloalkyl, substituted cycloalkyl, heterocycloalkyl,        substituted heterocycloalkyl, aryl, substituted aryl, arylalkyl,        substituted arylalkyl, heteroaryl, substituted heteroaryl,        heteroarylalkyl, and substituted heteroarylalkyl;    -   X₁ and X₂ are each independently selected from —C, —NR^(j), —O,        —SR^(k), —Si, wherein R^(j) and R^(k) are each independently        selected from hydrogen, alkyl or substituted alkyl, heteroalkyl,        substituted heteroalkyl, cycloalkyl, substituted cycloalkyl,        heterocycloalkyl, substituted heterocycloalkyl, aryl,        substituted aryl, arylalkyl, substituted arylalkyl, heteroaryl,        substituted heteroaryl, heteroarylalkyl, and substituted        heteroarylalkyl;    -   R^(a) is selected from hydrogen, oxygen, fluorine, —CF₃, or        together with X₁ form a cycloalkyl, substituted cycloalkyl,        heterocycloalkyl, substituted heterocycloalkyl, aryl,        substituted aryl, arylalkyl, substituted arylalkyl, heteroaryl,        substituted heteroaryl, heteroarylalkyl, and substituted        heteroarylalkyl;    -   wherein        indicates a double or single bond,    -   n is an integer from 2 to 25;    -   Y is selected from carbon, nitrogen or silicon;    -   R^(b), R^(c) and R^(d) are independently selected hydrogen,        alkyl, substituted alkyl, heteroalkyl, substituted heteroalkyl,        cycloalkyl, substituted cycloalkyl, heterocycloalkyl,        substituted heterocycloalkyl, aryl, substituted aryl, arylalkyl,        substituted arylalkyl, heteroaryl, substituted heteroaryl,        heteroarylalkyl, and substituted heteroarylalkyl, wherein when Y        is nitrogen, R^(d) is not present, or wherein R^(c) and R^(d)        together with Y form a cycloalkyl, substituted cycloalkyl, aryl,        substituted aryl, heteroaryl or substituted heteroaryl group;        and    -   R^(c) is alkyl or substituted alkyl,    -   or salt, solvate or hydrate thereof.

In some instances, R¹ is hydrogen. In some instances, R² is hydrogen. Insome instances, R³ is hydrogen. In some instances, R⁴ is hydrogen. Insome instances, each of R¹, R², R³, R⁴ are each hydrogen.

In certain instances, R¹ is:

-   -   wherein        represents the R¹—O bond;    -   R⁸ is hydrogen, alkyl or substituted alkyl; In some instances,        R⁸ is selected from hydrogen, methyl, ethyl and cyclopropyl.    -   R⁹ is —NR^(f) or —OR^(f), wherein R^(f) is alkyl, substituted        alkyl, acyl, alkylacyl or substituted alkylacyl, or wherein        R^(f) together with R¹⁰ form a cycloalkyl, substituted        cycloalkyl, heterocycloalkyl, substituted heterocycloalkyl,        aryl, substituted aryl, arylalkyl, substituted arylalkyl,        heteroaryl, substituted heteroaryl, heteroarylalkyl, and        substituted heteroarylalkyl; and    -   R¹⁰ is cycloalkyl, substituted cycloalkyl, heterocycloalkyl,        substituted heterocycloalkyl, aryl, substituted aryl, arylalkyl,        substituted arylalkyl, heteroaryl, substituted heteroaryl,        heteroarylalkyl, and substituted heteroarylalkyl or wherein R¹⁰        together with R^(f) form a cycloalkyl, substituted cycloalkyl,        heterocycloalkyl, substituted heterocycloalkyl, aryl,        substituted aryl, arylalkyl, substituted arylalkyl, heteroaryl,        substituted heteroaryl, heteroarylalkyl, and substituted        heteroarylalkyl.

In certain instances, R¹ is selected from:

-   -   wherein        represents the R¹—O bond.    -   R⁸ is hydrogen, alkyl, substituted alkyl; and    -   R^(g) is hydrogen or a halogen selected from fluorine, chlorine,        bromine or iodine.

In certain instances, R¹ is:

-   -   wherein:    -   R¹¹ is alkyl or substituted alkyl;    -   R¹² is alkyl or alkyl substituted with a cycloalkyl, substituted        cycloalkyl, heterocycloalkyl, substituted heterocycloalkyl,        aryl, substituted aryl, arylalkyl, substituted arylalkyl,        heteroaryl, substituted heteroaryl, heteroarylalkyl, and        substituted heteroarylalkyl.

In some instances, R¹¹ is selected from methyl, ethyl, n-propyl,iso-propyl, n-butyl, iso-butyl, t-butyl. In certain instances, R¹¹ isiso-propyl.

In certain instances, R¹ is:

-   -   wherein R^(h) and R^(i) are each independently selected from        hydrogen, hydroxyl or halogen. In some instances, R^(h) is        hydroxyl. In some instances, R^(h) is F. In some instances,        R^(h) is Cl. In some instances, R^(h) is I. In some instances,        R^(h) is Br. In some instances, R^(i) is hydroxyl. In some        instances, R^(i) is F. In other instances, R^(i) is Cl. In other        instances, R^(i) is I. In other instances, R^(i) is Br.

In some embodiments, X₁ is —NH. In some instances, X₁, X₂ and R^(a)together form an amide, such as where X₁ is —NH, X₂ is carbon and R^(a)is oxygen. In other instances, X₁, X₂ and R^(a) together form asulfoximine, such as where X₁ is —NH, X₂ is —SR and R^(a) is oxygen. Insome instances, R^(k) is selected from methyl, ethyl, n-propyl,iso-propyl, n-butyl, iso-butyl, t-butyl. In certain instances, R^(k) ismethyl. In other instances, X₁, X₂ and R^(a) together form atrifluoromethyl aminomethyl, such as where X₁ is —NH, X₂ is carbon andR^(a) is trifluoromethyl. In other instances, X₁, X₂ and R^(a) togetherform a vinylfluoride, such as where X₁ is carbon. X₂ is carbon and R^(a)is fluorine. In certain instances, X₁, X₂ and R^(a) together form anaminooxetan, such as where X₁ is —NH and R³ together with X₂ forms anoxacyclobutane.

In embodiments. R^(c) may be alkyl or substituted alkyl. In someinstances, R^(e) is a C8 to C20 alkyl. In some instances, R^(e) is asubstituted C8 to C20 alkyl. In certain instances, R^(e) is a C13 alkyl.

In some embodiments, R^(b) is hydrogen. In some instances, R^(b) is a C1to C16 alkyl. In some instances, R^(a) is selected from the groupconsisting of methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl,tert-butyl, pentyl, hexyl, heptyl and octyl. In some instances, R^(b) isselected from:

-   -   wherein        indicates a bond to Y and R^(z) is hydrogen, alkyl or alkyl        substituted with a cycloalkyl, substituted cycloalkyl,        heterocycloalkyl, substituted heterocycloalkyl, aryl,        substituted aryl, arylalkyl, substituted arylalkyl, heteroaryl,        substituted heteroaryl, heteroarylalkyl, and substituted        heteroarylalkyl. In certain instances, W is alkyl, such as a C1        to C16 alkyl or C1 to C16 substituted alkyl.

In some embodiments, R^(c) is a C1 to C16 alkyl. In some instances,R^(c) is selected from methyl, ethyl, propyl, isopropyl, n-butyl,isobutyl, tert-butyl, pentyl, hexyl, heptyl and octyl.

In some embodiments, R^(d) is a C5 to C25 alkyl or a C5 to C25 alkylsubstituted with a cycloalkyl group, substituted cycloalkyl group,heterocycloalkyl group, substituted heterocycloalkyl group, aryl group,substituted aryl group, arylalkyl group, substituted arylalkyl group,heteroaryl group, substituted heteroaryl group, heteroarylalkyl group,or substituted heteroarylalkyl group. In some instances, R^(d) is a C5to C25 alkyl substituted with a moiety selected from the groupconsisting of:

-   -   wherein        indicates the bond to the C5 to C25 alkyl, and R^(m) and R^(n)        are independently selected from hydrogen, halogen, hydroxyl,        substituted hydroxyl, amino, substituted amino, thiol,        substituted thiol, sulfoxide, substituted sulfoxide, sulfone,        substituted sulfone, sulfoximine, substituted sulfoximine, Acyl,        aminoacyl, alkyl, substituted alkyl; heteroalkyl, substituted        heteroalkyl, cycloalkyl, substituted cycloalkyl, spiroalkyl,        heterocycloalkyl, substituted heterocycloalkyl, aryl,        substituted aryl, arylalkyl, substituted arylalkyl, heteroaryl,        substituted heteroaryl, heteroarylalkyl, and substituted        heteroarylalkyl.

In some instances, R^(m) is hydrogen. In some instances, R^(m) ishalogen. In some instances, R^(m) is selected from fluorine, bromine oriodine. In some instances, R^(n) is hydrogen. In some instances, R^(n)is halogen. In some instances, R^(n) is fluorine, bromine or iodine.

In some embodiments, compounds of interest include a compound of formulaDCD-(II):

-   -   wherein:    -   Z is selected from:

-   -   wherein        indicates the Z—O bond;    -   wherein X is —NHCO— or oxygen    -   R¹, R², R³ and R⁴ are each independently selected from hydrogen,        alkyl, substituted alkyl, heteroalkyl, substituted heteroalkyl,        cycloalkyl, substituted cycloalkyl, heterocycloalkyl,        substituted heterocycloalkyl, aryl, substituted aryl, arylalkyl,        substituted arylalkyl, heteroaryl, substituted heteroaryl,        heteroarylalkyl, and substituted heteroarylalkyl;    -   X₁, X₂, X₃, X₄ and X₅ are each independently selected from        carbon, nitrogen, oxygen or sulfur;    -   R^(a) is optionally absent or when present is selected from        hydrogen or oxygen;    -   wherein        indicates a double or single bond;    -   n is an integer from 2 to 25;    -   Y is selected from carbon, nitrogen or silicon;    -   R^(b), R^(c) and R^(d) are independently selected hydrogen,        alkyl, substituted alkyl, heteroalkyl, substituted heteroalkyl,        cycloalkyl, substituted cycloalkyl, heterocycloalkyl,        substituted heterocycloalkyl, aryl, substituted aryl, arylalkyl,        substituted arylalkyl, heteroaryl, substituted heteroaryl,        heteroarylalkyl, and substituted heteroarylalkyl, wherein when Y        is nitrogen, R^(d) is not present, or wherein R^(c) and R^(d)        together with Y form a cycloalkyl, substituted cycloalkyl, aryl,        substituted aryl, heteroaryl or substituted heteroaryl group;        and    -   R^(e) is alkyl or substituted alkyl,    -   or salt, solvate or hydrate thereof.

In some instances, R¹ is hydrogen. In some instances, R² is hydrogen. Insome instances. R³ is hydrogen. In some instances, R⁴ is hydrogen. Insome instances, each of R¹, R², R³, R⁴ are each hydrogen.

In certain instances, R¹ is:

-   -   wherein        represents the R¹—O bond;    -   R⁸ is hydrogen, alkyl or substituted alkyl; In some instances,        R⁸ is selected from hydrogen, methyl, ethyl and cyclopropyl.    -   R⁹ is —NR^(f) or —OR^(f), wherein R^(f) is alkyl, substituted        alkyl, acyl, alkylacyl or substituted alkylacyl, or wherein        R^(f) together with R¹⁰ form a cycloalkyl, substituted        cycloalkyl, heterocycloalkyl, substituted heterocycloalkyl,        aryl, substituted aryl, arylalkyl, substituted arylalkyl,        heteroaryl, substituted heteroaryl, heteroarylalkyl, and        substituted heteroarylalkyl; and    -   R¹⁰ is cycloalkyl, substituted cycloalkyl, heterocycloalkyl,        substituted heterocycloalkyl, aryl, substituted aryl, arylalkyl,        substituted arylalkyl, heteroaryl, substituted heteroaryl,        heteroarylalkyl, and substituted heteroarylalkyl or wherein R¹⁰        together with R^(f) form a cycloalkyl, substituted cycloalkyl,        heterocycloalkyl, substituted heterocycloalkyl, aryl,        substituted aryl, arylalkyl, substituted arylalkyl, heteroaryl,        substituted heteroaryl, heteroarylalkyl, and substituted        heteroarylalkyl.

In certain instances, R¹ is selected from:

-   -   wherein        represents the R¹—O bond;    -   R⁸ is hydrogen, alkyl, substituted alkyl; and    -   R^(g) is hydrogen or a halogen selected from fluorine, chlorine,        bromine or iodine.

In certain instances, R¹ is:

-   -   wherein:    -   R¹¹ is alkyl or substituted alkyl;    -   R¹² is alkyl or alkyl substituted with a cycloalkyl, substituted        cycloalkyl, heterocycloalkyl, substituted heterocycloalkyl,        aryl, substituted aryl, arylalkyl, substituted arylalkyl,        heteroaryl, substituted heteroaryl, heteroarylalkyl, and        substituted heteroarylalkyl.

In some instances, R¹¹ is selected from methyl, ethyl, n-propyl,iso-propyl, n-butyl, iso-butyl, t-butyl. In certain instances, R¹¹ isiso-propyl.

In certain instances, R¹ is:

-   -   wherein R^(h) and R^(i) are each independently selected from        hydrogen, hydroxyl or halogen. In some instances, R^(h) is        hydroxyl. In some instances, R^(h) is F. In some instances,        R^(h) is Cl. In some instances, R^(h) is I. In some instances,        R^(i) is Br. In some instances, R^(i) is hydroxyl. In some        instances, R^(i) is F. In other instances, R^(i) is Cl. In other        instances, R^(i) is I. In other instances, R^(i) is Br.

In some embodiments, X₁, X₂, X₃, X₄ and X₅ together form a pyrazole,such as where X₁ is carbon, X₂ is nitrogen, X₃ is nitrogen, X₄ is carbonand X₅ is carbon. In other embodiments, X₁, X₂, X₃, X₄ and X₅ togetherform an imidazole, such as where X₁ is carbon, X₂ is nitrogen, X₃ iscarbon, X₄ is carbon and X₅ is nitrogen. In other embodiments, X₁, X₂,X₃, X₄ and X₅ together form a tetrazole, such as where X₁ is nitrogen,X₂ is nitrogen, X₃ is nitrogen, X₄ is nitrogen and X₅ is carbon. Incertain embodiments, X₁, X₂, X₃, X₄ and X₅ together form a tetrazoline,such as where X₁ is nitrogen, X₂ is nitrogen, X₃ is nitrogen, X₄ isnitrogen, X₅ is carbon and R^(a) is oxygen.

In embodiments, R^(e) may be alkyl or substituted alkyl. In someinstances, R^(e) is a C8 to C20 alkyl. In some instances, R^(e) is asubstituted C5 to C20 alkyl. In certain instances, R^(e) is a C13 alkyl.

In some embodiments, R^(b) is hydrogen. In some instances, R^(b) is a C1to C16 alkyl. In some instances, R^(b) is selected from the groupconsisting of methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl,tert-butyl, pentyl, hexyl, heptyl and octyl. In some instances, R^(a) isselected from.

-   -   wherein        indicates a bond to Y and R^(z) is hydrogen, alkyl or alkyl        substituted with a cycloalkyl, substituted cycloalkyl,        heterocycloalkyl, substituted heterocycloalkyl, aryl,        substituted aryl, arylalkyl, substituted arylalkyl, heteroaryl,        substituted heteroaryl, heteroarylalkyl, and substituted        heteroarylalkyl. In certain instances, R^(z) is alkyl, such as a        C1 to C16 alkyl or C1 to C16 substituted alkyl.

In some embodiments, R^(c) is a C1 to C16 alkyl. In some instances,R^(c) is selected from the group consisting of methyl, ethyl, propyl,isopropyl, n-butyl, isobutyl, tert-butyl, pentyl, hexyl, heptyl andoctyl.

In some embodiments, R^(d) is a C5 to C25 alkyl or a C5 to C25 alkylsubstituted with a cycloalkyl group, substituted cycloalkyl group,heterocycloalkyl group, substituted heterocycloalkyl group, aryl group,substituted aryl group, arylalkyl group, substituted arylalkyl group,heteroaryl group, substituted heteroaryl group, heteroarylalkyl group,or substituted heteroarylalkyl group. In some instances, R^(d) is a C5to C25 alkyl substituted with a moiety selected from the groupconsisting of:

-   -   wherein        indicates the bond to the C5 to C25 alkyl; and R^(m) and R^(n)        are independently selected from hydrogen, halogen, hydroxyl,        substituted hydroxyl, amino, substituted amino, thiol,        substituted thiol, sulfoxide, substituted sulfoxide, sulfone,        substituted sulfone, sulfoximine, substituted sulfoximine. Acyl,        aminoacyl, alkyl, substituted alkyl, heteroalkyl, substituted        heteroalkyl, cycloalkyl, substituted cycloalkyl, spiroalkyl,        heterocycloalkyl, substituted heterocycloalkyl, aryl,        substituted aryl, arylalkyl, substituted arylalkyl, heteroaryl,        substituted heteroaryl, heteroarylalkyl, and substituted        heteroarylalkyl.

In some instances, R^(m) is hydrogen. In some instances, R^(m) ishalogen. In some instances, R^(m) is selected from fluorine, bromine oriodine. In some instances, R^(n) is hydrogen. In some instances, R^(n)is halogen. In some instances, R^(n) is fluorine, bromine or iodine.

Aspects of the disclosure also include methods for activing an iNKT cellby contacting the iNKT cell with an amount of the subject compounds or apharmaceutically acceptable salt thereof. In some instances, the iNKTcell is contacted with the compound in vitro. In other instances, theiNKT cell is contacted with the compound in vivo. In some instances,methods include contacting one or more of the compounds described hereinwith iNKT cells in a manner sufficient to induce a T_(H)1-type cytokineresponse (e.g., increase production of one or more cytokines selectedfrom IFNγ, IL-1β, IL-2, IL-3, IL-8, IL-12, IL-15, TNFα, GM-CSF, RANTES,MIP-1α and MCP-1). In other instances, methods include contacting one ormore of the compounds described herein with iNKT cells in a mannersufficient to induce a T_(H)2-type cytokine response (e.g., increaseproduction of one or more cytokines selected from IL-4, IL-6, IL-8,IL-10, IL-13, RANTES, MIP-1α and MCP-1) In some instances, the compoundforms a complex with a CD1 molecule on an antigen-presenting cell. Incertain instances, the CD1 molecule is a CD1d molecule. In someinstances, the receptor on the T lymphocyte is a T cell receptor. Insome instances, the compound stimulates at least one other lymphocyte toproduce the cytokine response in some instances the at least one otherlymphocyte is a T helper cell.

In certain instances, methods include contacting activated iNKT cellswith a composition comprising senescent cells where contacting theactivated iNKT cells reduces the presence of or eliminates the senescentcells in the composition. In some embodiments, the senescent cells havean inflammatory secretome. In some embodiments, the composition furtherincludes healthy cells. In some instances, contacting the activated iNKTcells reduces the presence of or eliminates the senescent cells in thecomposition without reducing the presence of the healthy cells. Forexample, contacting the activated iNKT cells reduces the presence of oreliminates the senescent cells in the composition and the presence ofhealthy cells is reduced by 5% or less when the composition is contactedwith the activated iNKT cells.

In some embodiments, methods include administering one or more of thecompounds described herein to a subject, such as to reduce the presenceof or eliminate senescent cells in the subject. In some instances,methods include administering one or more of the compounds to treat thesubject for an autoimmune disease, fibrotic disorders (lung, kidney,liver), an allergic disease, a metabolic syndrome, type 2 diabetes,NAFLD, NASH, cancer, an eye disease, heart disease, kidney disease,pathogen infection, rheumatoid arthritis, ulcerative colitis, multiplesclerosis, familial hypercholesteremia, giant cell arteritis, idiopathicpulmonary fibrosis, systemic lupus erythematosus, cachexia, glaucoma,chronic obstructive pulmonary disease, systemic sclerosis, pulmonaryarterial hypertension, lipodystrophy, sarcopenia, alopecia, postmyocardial infarction, vitiligo, POTS, MCAD, Sjogren's, Scleroderma,Hashimoto Disease, Ankylosing Spondylitis, Fibromyalgia, Sarcoidosis.Hepatitis, Raynauld's Syndrome, Mold Illness, Celiac, Crohn's,Pemphigus. SPS, PBC, Psoriatic Arthritis, CIDP, motor neuron disease,GPA, ALS, myasenthia gravis, and presbyopia.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1A depict the percentage GFP+ of cells at different concentrationsof compounds DCD-101, DCD-102, DCD-103, DCD-104, DCD-105, DCD-106,DCD-108, DCD-112, DCD-113, DCD-114, DCD115, and DCD-116, DCD118,DCD-119, DCD-120, DCD-121, DCD-122, DCD-123, DCD-124, DCD-125, DCD-126,DCD-127, DCD-128, DCD-129, DCD-130, DCD-131, DCD-132, DCD-133, DCD-134,DCD-135, DCD-136, DCD-137, DCD-138, DCD-139, DCD-140, DCD-141, DCD-142,DCD-143, DCD-144, DCD-145, DCD-146, DCD-147, DCD-148, DCD-149, DCD-150,DCD-151, DCD-152, DCD-153, DCD-154, DCD-155, DCD-156, DCD-157, DCD-158,DCD-159 and α-GalCer at varying concentrations.

FIG. 1B depict the EC50 of the dose responses shown in FIG. 1A.

FIG. 2A depicts the amount of interleukin-2 (IL-2) secretion in responseto a 48 hour incubation with compounds DCD-101, DCD-102, DCD-104,DCD-105, DCD-106 and α-GalCer in a dose response of 0.01, 0.1, 1, and 10ug/mL.

FIG. 3A depicts the secretion of the cytokine interferon gamma (IFNγ) inresponse to activation by compounds DCD-101, DCD-104, DCD-106 andα-GalCer.

FIG. 3B depicts the secretion of the cytokine interleukin-6 (IL-6) inresponse to activation by compounds DCD-101, DCD-104, DCD-106 andα-GalCer.

FIG. 3C depicts the secretion of the cytokine tumor necrosis factoralpha (TNFα) in response to activation by compounds DCD-101, DCD-104,DCD-106 and α-GalCer.

FIG. 4A depicts the activation of C57BL/6J immune cells, as measured bythe serum IFNγ using ELISA, in response to injection of the compoundsDCD-101, DCD-119, DCD-123, DCD125, DCD127, DCD-128, DCD-134, DCD-142,DCD-145, DCD-146, DCD-147, DCD-148, DCD-149, DCD-150, DCD-151, DCD-152,DCD-153, DCD-154, DCD-155, DCD-156, and DCD-157, DCD-158, and DCD-159with comparison to αGalCer twenty hours after injection.

FIG. 4B depicts the expansion of iNKT cells in the C57BL/6J mouse spleenin response to injection of the compounds DCD-101, DCD-104, DCD-106,DCD-119, DCD-142, DCD-145, DCD-146, DCD-147, DCD-148, DCD-149, DCD-150,DCD-151, DCD-152, DCD-153, DCD-154, DCD-155, DCD-156, DCD-157, DCD-158,and DCD-159 with comparison to αGalCer.

FIG. 5A depicts the expansion of iNKT cells caused by αGalCer or DCD-101in a HFD model mouse spleen, as measured by flow cytometry.

FIG. 5B depicts the expansion of iNKT cells caused by αGalCer or DCD-154in the HFD mouse model eWAT, as measured by flow cytometry.

FIG. 5C depicts the amount of IFNγ in blood serum generated in the HFDmouse model in response to αGalCer or DCD-101 two hours after injection.

FIG. 5D depicts the amount of IFNγ in blood serum generated in the HFDmouse model in response to αGalCer or DCD-154.20 hours after injection.

FIG. 5E depicts the reduction in accumulated senescent cells in eWAT inresponse to treatment with compound DCD-101, DCD-154 and αGalCer. Valueswere collected from multiple experiments and normalized to theHFD-vehicle condition.

FIG. 6A depicts expression of GFP in response to incubation withcompound GVK1a, GVK1b, GVK1c, and GVK1f with BWSTIM cells and JiNKTcells.

FIG. 6B depicts IL-2 expression by compounds GVK1a, GVK1b, and GVK1f inthe DN3.2 cell line when loaded on BWSTIM CD1d.

FIG. 6C depicts the secretion of the cytokine interferon gamma (IFNγ),tumor necrosis factor alpha (TNFα), interleukin-4 (IL-4) andinterleukin-6 (IL-6) in response to incubation with compounds GVK1a,GVK1b, GVK1c and GVK1f and α-GalCer.

FIG. 7A depicts the effect of activated iNKT cells on senescent cellsand non-senescent cells over a period of 18 hours.

FIG. 7B depicts a comparison between cytolysis of senescent cells andhealthy cells by activated iNKT cells after incubation for 8 hours and18 hours.

DEFINITIONS

The following terms have the following meaning unless otherwiseindicated. Any undefined terms have their art recognized meanings.

As used herein, the term “alkyl” by itself or as part of anothersubstituent refers to a saturated branched or straight-chain monovalenthydrocarbon radical derived by the removal of one hydrogen atom from asingle carbon atom of a parent alkane. Typical alkyl groups include, butare not limited to, methyl; ethyl, propyls such as propan-1-yl orpropan-2-yl; and butyls such as butan-1-yl, butan-2-yl,2-methyl-propan-1-yl or 2-methyl-propan-2-yl. In some embodiments, analkyl group comprises from 1 to 20 carbon atoms. In other embodiments,an alkyl group comprises from 1 to 10 carbon atoms. In still otherembodiments, an alkyl group comprises from 1 to 6 carbon atoms, such asfrom 1 to 4 carbon atoms.

“Alkanyl” by itself or as part of another substituent refers to asaturated branched, straight-chain or cyclic alkyl radical derived bythe removal of one hydrogen atom from a single carbon atom of an alkane.Typical alkanyl groups include, but are not limited to, methanyl;ethanyl; propanyls such as propan-1-yl, propan-2-yl (isopropyl),aggregate-1-yl, etc.; butanyls such as butan-1-yl, butan-2-yl(sec-butyl), 2-methyl-propan-1-yl (isobutyl), 2-methyl-propan-2-yl(t-butyl), cyclobutan-1-yl, etc.; and the like.

“Alkylene” refers to a branched or unbranched saturated hydrocarbonchain, usually having from 1 to 40 carbon atoms, more usually 1 to 10carbon atoms and even more usually 1 to 6 carbon atoms. This term isexemplified by groups such as methylene (—CH₂—), ethylene (—CH₂CH₂—),the propylene isomers (e.g., —CH₂CH₂CH₂— and —CH(CH₃)CH₂—) and the like.

“Alkenyl” by itself or as part of another substituent refers to anunsaturated branched, straight-chain or cyclic alkyl radical having atleast one carbon-carbon double bond derived by the removal of onehydrogen atom from a single carbon atom of an alkene. The group may bein either the cis or trans conformation about the double bond(s).Typical alkenyl groups include, but are not limited to, ethenyl;propenyls such as prop-1-en-1-yl, prop-1-en-2-yl, prop-2-en-1-yl(allyl), prop-2-en-2-yl, cycloprop-1-en-1-yl; cycloprop-2-en-1-yl;butenyls such as but-1-en-1-yl, but-1-en-2-yl, 2-methyl-prop-1-en-1-yl,but-2-en-1-yl, but-2-en-1-yl, but-2-en-2-yl, buta-1,3-dien-1-yl,buta-1,3-dien-2-yl, cyclobut-1-en-1-yl, cyclobut-1-en-3-yl,cyclobuta-1,3-dien-1-yl, etc.; and the like.

“Alkynyl” by itself or as part of another substituent refers to anunsaturated branched, straight-chain or cyclic alkyl radical having atleast one carbon-carbon triple bond derived by the removal of onehydrogen atom from a single carbon atom of an alkyne. Typical alkynylgroups include, but are not limited to, ethynyl; propynyls such asprop-1-yn-1-yl, prop-2-yn-1-yl, etc., butynyls such as but-1-yn-1-yl,but-1-yn-3-yl, but-3-yn-1-yl, etc.; and the like.

“Acyl” by itself or as part of another substituent refers to a radical—C(O)R³⁰, where R³⁰ is hydrogen, alkyl, cycloalkyl, cycloheteroalkyl,aryl, arylalkyl, heteroalkyl, heteroaryl, heteroarylalkyl as definedherein and substituted versions thereof. Representative examplesinclude, but are not limited to formyl, acetyl, cyclohexylcarbonyl,cyclohexylmethylcarbonyl, benzoyl, benzylcarbonyl, piperonyl, succinyl,and malonyl, and the like.

The term “aminoacyl” refers to the group —C(O)NR²¹R²², wherein R²¹ andR²² independently are selected from the group consisting of hydrogen,alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl,substituted alkynyl, aryl, substituted aryl, cycloalkyl, substitutedcycloalkyl, cycloalkenyl, substituted cycloalkenyl, heteroaryl,substituted heteroaryl, heterocyclic, and substituted heterocyclic andwhere R²¹ and R²² are optionally joined together with the nitrogen boundthereto to form a heterocyclic or substituted heterocyclic group, andwherein alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl,substituted alkynyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl,substituted cycloalkenyl, aryl, substituted aryl, heteroaryl,substituted heteroaryl, heterocyclic, and substituted heterocyclic areas defined herein.

“Alkoxy” by itself or as part of another substituent refers to a radical—OR³¹ where R³¹ represents an alkyl or cycloalkyl group as definedherein. Representative examples include, but are not limited to,methoxy, ethoxy, propoxy, butoxy, cyclohexyloxy and the like.

“Alkoxycarbonyl” by itself or as part of another substituent refers to aradical —C(O)OR³¹ where R³¹ represents an alkyl or cycloalkyl group asdefined herein. Representative examples include, but are not limited to,methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, butoxycarbonyl,cyclohexyloxycarbonyl and the like.

“Aryl” by itself or as part of another substituent refers to amonovalent aromatic hydrocarbon radical derived by the removal of onehydrogen atom from a single carbon atom of an aromatic ring system.Typical aryl groups include, but are not limited to, groups derived fromaceanthrylene, acenaphthylene, acephenanthrylene, anthracene, azulene,benzene, chrysene, coronene, fluoranthene, fluorene, hexacene,hexaphene, hexalene, as-indacene, s-indacene, indane, indene,naphthalene, octacene, octaphene, octalene, ovalene, penta-2,4-diene,pentacene, pentalene, pentaphene, perylene, phenalene, phenanthrene,picene, pleiadene, pyrene, pyranthrene, rubicene, triphenylene,trinaphthalene and the like. In certain embodiments, an aryl groupcomprises from 6 to 20 carbon atoms. In certain embodiments, an arylgroup comprises from 6 to 12 carbon atoms. Examples of an aryl group arephenyl and naphthyl.

“Arylalkyl” by itself or as part of another substituent refers to anacyclic alkyl radical in which one of the hydrogen atoms bonded to acarbon atom, typically a terminal or sp³ carbon atom, is replaced withan aryl group. Typical arylalkyl groups include, but are not limited to,benzyl, 2-phenylethan-1-yl, 2-phenylethen-1-yl, naphthylmethyl,2-naphthylethan-1-yl, 2-naphthylethen-1-yl, naphthobenzyl,2-naphthophenylethan-1-yl and the like. Where specific alkyl moietiesare intended, the nomenclature arylalkanyl, arylalkenyl and/orarylalkynyl is used. In certain embodiments, an arylalkyl group is(C₇-C₃₀) arylalkyl, e.g., the alkanyl, alkenyl or alkynyl moiety of thearylalkyl group is (C₁-C₁₀) and the aryl moiety is (C₆-C₂₀). In certainembodiments, an arylalkyl group is (C₇-C₂₀) arylalkyl, e.g., thealkanyl, alkenyl or alkynyl moiety of the arylalkyl group is (C₁-C₈) andthe aryl moiety is (C₆-C₁₂).

“Arylaryl” by itself or as part of another substituent, refers to amonovalent hydrocarbon group derived by the removal of one hydrogen atomfrom a single carbon atom of a ring system in which two or moreidentical or non-identical aromatic ring systems are joined directlytogether by a single bond, where the number of such direct ringjunctions is one less than the number of aromatic ring systems involved.Typical arylaryl groups include, but are not limited to, biphenyl,triphenyl, phenyl-napthyl, binaphthyl, biphenyl-napthyl, and the like.When the number of carbon atoms in an arylaryl group are specified, thenumbers refer to the carbon atoms comprising each aromatic ring. Forexample, (C₅-C₁₄) arylaryl is an arylaryl group in which each aromaticring comprises from 5 to 14 carbons, e.g., biphenyl, triphenyl,binaphthyl, phenylnapthyl, etc. In certain embodiments, each aromaticring system of an arylaryl group is independently a (C₅-C₁₄) aromatic.In certain embodiments, each aromatic ring system of an arylaryl groupis independently a (C₅-C₁₀) aromatic. In certain embodiments, eacharomatic ring system is identical. e.g., biphenyl, triphenyl,binaphthyl, trinaphthyl, etc.

“Cycloalkyl” by itself or as part of another substituent refers to asaturated or unsaturated cyclic alkyl radical. Where a specific level ofsaturation is intended, the nomenclature “cycloalkanyl” or“cycloalkenyl” is used. Typical cycloalkyl groups include, but are notlimited to, groups derived from cyclopropane, aggregate e, cyclopentane,cyclohexane and the like. In certain embodiments, the cycloalkyl groupis (C₃-C₁₀) cycloalkyl. In certain embodiments, the cycloalkyl group is(C₃-C₇) cycloalkyl.

“Cycloheteroalkyl” or “heterocyclyl” by itself or as part of anothersubstituent, refers to a saturated or unsaturated cyclic alkyl radicalin which one or more carbon atoms (and any associated hydrogen atoms)are independently replaced with the same or different heteroatom.Typical heteroatoms to replace the carbon atom(s) include, but are notlimited to, N, P, O, S, Si, etc. Where a specific level of saturation isintended, the nomenclature “cycloheteroalkanyl” or “cycloheteroalkenyl”is used. Typical cycloheteroalkyl groups include, but are not limitedto, groups derived from epoxides, azirines, thiiranes, imidazolidine,morpholine, piperazine, piperidine, pyrazolidine, pyrrolidine,quinuclidine and the like.

“Heteroalkyl, Heteroalkanyl, Heteroalkenyl and Heteroalkynyl” bythemselves or as part of another substituent refer to alkyl, alkanyl,alkenyl and alkynyl groups, respectively, in which one or more of thecarbon atoms (and any associated hydrogen atoms) are independentlyreplaced with the same or different heteroatomic groups. Typicalheteroatomic groups which can be included in these groups include, butare not limited to, —O—, —S—, —S—S—, —O—S—, —NR³⁷R³⁸—, ═N—N═, —N═N—,—N═N—NR³⁹R⁴⁰, —PR⁴¹—, —P(O)₂—, —POR⁴²—, —O—P(O)₂—, —S—O—, —S—(O)—,—SO₂—, —SnR⁴³R⁴⁴— and the like, where R³⁷, R³⁸, R³⁹, R⁴⁰, R⁴¹, R⁴², R⁴³and R⁴⁴ are independently hydrogen, alkyl, substituted alkyl, aryl,substituted aryl, arylalkyl, substituted arylalkyl, cycloalkyl,substituted cycloalkyl, cycloheteroalkyl, substituted cycloheteroalkyl,heteroalkyl, substituted heteroalkyl, heteroaryl, substitutedheteroaryl, heteroarylalkyl or substituted heteroarylalkyl.

“Heteroaryl” by itself or as part of another substituent, refers to amonovalent heteroaromatic radical derived by the removal of one hydrogenatom from a single atom of a heteroaromatic ring system. Typicalheteroaryl groups include, but are not limited to, groups derived fromacridine, arsindole, carbazole, β-carboline, chromane, chromene,cinnoline, furan, imidazole, indazole, indole, indoline, indolizine,isobenzofuran, isochromene, isoindole, isoindoline, isoquinoline,isothiazole, isoxazole, naphthyndine, oxadiazole, oxazole, perimidine,phenanthridine, phenanthroline, phenazine, phthalazine, pteridine,purine, pyran, pyrazine, pyrazole, pyridazine, pyridine, pyrimidine,pyrrole, pyrrolizine, quinazoline, quinoline, quinolizine, quinoxaline,tetrazole, thiadiazole, thiazole, thiophene, triazole, xanthene,benzodioxole and the like. In certain embodiments, the heteroaryl groupis from 5-20 membered heteroaryl. In certain embodiments, the heteroarylgroup is from 5-10 membered heteroaryl. In certain embodiments,heteroaryl groups are those derived from thiophene, pyrrole,benzothiophene, benzofuran, indole, pyridine, quinoline, imidazole,oxazole and pyrazine.

“Heteroarylalkyl” by itself or as part of another substituent, refers toan acyclic alkyl radical in which one of the hydrogen atoms bonded to acarbon atom, typically a terminal or sp³ carbon atom, is replaced with aheteroaryl group. Where specific alkyl moieties are intended, thenomenclature heteroarylalkanyl, heteroarylalkenyl and/orheterorylalkynyl is used. In certain embodiments, the heteroarylalkylgroup is a 6-30 membered heteroarylalkyl, e.g., the alkanyl, alkenyl oralkynyl moiety of the heteroarylalkyl is 1-10 membered and theheteroaryl moiety is a 5-20-membered heteroaryl. In certain embodiments,the heteroarylalkyl group is 6-20 membered heteroarylalkyl, e.g., thealkanyl, alkenyl or alkynyl moiety of the heteroarylalkyl is 1-8membered and the heteroaryl moiety is a 5-12-membered heteroaryl.

“Aromatic Ring System” by itself or as part of another substituent,refers to an unsaturated cyclic or polycyclic ring system having aconjugated π electron system. Specifically included within thedefinition of “aromatic ring system” are fused ring systems in which oneor more of the rings are aromatic and one or more of the rings aresaturated or unsaturated, such as, for example, fluorene, indane,indene, phenalene, etc. Typical aromatic ring systems include, but arenot limited to, aceanthrylene, acenaphthylene, acephenanthrylene,anthracene, azulene, benzene, chrysene, coronene, fluoranthene,fluorene, hexacene, hexaphene, hexalene, as-indacene, s-indacene,indane, indene, naphthalene, octacene, octaphene, octalene, ovalene,penta-2,4-diene, pentacene, pentalene, pentaphene, perylene, phenalene,phenanthrene, picene, pleiadene, pyrene, pyranthrene, rubicene,triphenylene, trinaphthalene and the like.

“Heteroaromatic Ring System” by itself or as part of anothersubstituent, refers to an aromatic ring system in which one or morecarbon atoms (and any associated hydrogen atoms) are independentlyreplaced with the same or different heteroatom. Typical heteroatoms toreplace the carbon atoms include, but are not limited to, N, P, O, S,Si, etc. Specifically included within the definition of “heteroaromaticring systems” are fused ring systems in which one or more of the ringsare aromatic and one or more of the rings are saturated or unsaturated,such as, for example, arsindole, benzodioxan, benzofuran, chromane,chromene, indole, indoline, xanthene, etc. Typical heteroaromatic ringsystems include, but are not limited to, arsindole, carbazole,β-carboline, chromane, chromene, cinnoline, furan, imidazole, indazole,indole, indoline, indolizine, isobenzofuran, isochromene, isoindole,isoindoline, isoquinoline, isothiazole, isoxazole, naphthyridine,oxadiazole, oxazole, perimidine, phenanthridine, phenanthroline,phenazine, phthalazine, pteridine, purine, pyran, pyrazine, pyrazole,pyridazine, pyridine, pyrimidine, pyrrole, pyrrolizine, quinazoline,quinoline, quinolizine, quinoxaline, tetrazole, thiadiazole, thiazole,thiophene, triazole, xanthene and the like.

“Substituted” refers to a group in which one or more hydrogen atoms areindependently replaced with the same or different substituent(s).Typical substituents include, but are not limited to, alkylenedioxy(such as methylenedioxy), -M, —R⁶⁰, —O—, ═O, —OR⁶⁰, —SR⁶⁰, —S⁻, ═S,—NR⁶⁰R⁶¹, —NR⁶⁰, —CF₃, —CN, —OCN, —SCN, —NO, —NO₂, ═N₂, —N₃, —S(O)₂O—,—S(O)₂OH, —S(O)₂R⁶⁰, —OS(O)₂O—, —OS(O)₂R⁶⁰, —P(O)(O⁻)₂, —P(O)(OR⁶⁰)(O⁻),—OP(O)(OR⁶⁰)(OR⁶¹), —C(O)R⁶⁰, —C(S)R⁶⁰, —C(O)OR⁶⁰, —C(O)NR⁶⁰R⁶¹,—C(O)O—, —C(S)OR⁶⁰, —NR⁶²C(O)NR⁶⁰R⁶¹, —NR⁶²C(S)NR⁶⁰R⁶¹,—NR⁶²C(NR⁶³)NR⁶⁰R⁶¹ and —C(NR⁶²)NR⁶⁰R⁶¹ where M is halogen; R⁶⁰, R⁶¹,R⁶² and R⁶³ are independently hydrogen, alkyl, substituted alkyl,alkoxy, substituted alkoxy, cycloalkyl, substituted cycloalkyl,cycloheteroalkyl, substituted cycloheteroalkyl, aryl, substituted aryl,heteroaryl or substituted heteroaryl, or optionally R⁶⁰ and R⁶¹ togetherwith the nitrogen atom to which they are bonded form a cycloheteroalkylor substituted cycloheteroalkyl ring; and R⁶⁴ and R⁶⁵ are independentlyhydrogen, alkyl, substituted alkyl, aryl, cycloalkyl, substitutedcycloalkyl, cycloheteroalkyl, substituted cycloheteroalkyl, aryl,substituted aryl, heteroaryl or substituted heteroaryl, or optionallyR⁶⁴ and R⁶⁵ together with the nitrogen atom to which they are bondedform a cycloheteroalkyl or substituted cycloheteroalkyl ring. In certainembodiments, substituents include -M, —R⁶⁰, ═O, —OR⁶⁰, —SR⁶⁰, —S⁻, ═S,—NR⁶⁰R⁶¹, ═NR⁶⁰, —CF₃, —CN, —OCN, —SCN, —NO, —NO₂, ═N₂, —N₃, —S(O)₂R⁶⁰,—OS(O)₂ ⁰⁻, —OS(O)₂R⁶⁰, —P(O)(O⁻)₂, —P(O)(OR(O⁻), —OP(O)(OR⁶⁰)(OR⁶¹),—C(O)R⁶⁰, —C(S)R⁶⁰, —C(O)OR⁶⁰, —C(O)NR⁶⁰R⁶¹, —C(O)O—, —NR⁶⁰,C(O)NR⁶⁰R⁶¹. In certain embodiments, substituents include -M, —R⁶⁰, ═O,—OR⁶⁰, —SR⁶⁰, —NR⁶⁰R⁶¹, —CF₃, —CN, —NO₂, —S(O)₂R⁶⁰, —P(O)(OR⁶⁰)(O⁻),—OP(O)(OR⁶⁰)(OR⁶¹), —C(O)R⁶⁰, —C(O)OR⁶⁰, —C(O)NR⁶⁰R⁶¹, —C(O)O⁻. Incertain embodiments, substituents include -M, —R⁶⁰, ═O, —OR⁶⁰, —SR⁶⁰,—NR⁶⁰R⁶¹, —CF₃, —CN, —NO₂, —S(O)₂R⁶⁰, —OP(O)(OR⁶⁰)OR), —C(O)R⁶⁰,—C(O)OR⁶⁰, —C(O)O⁻, where R⁶⁰, R⁶¹ and R⁶² are as defined above. Forexample, a substituted group may bear a methylenedioxy substituent orone, two, or three substituents selected from a halogen atom, a(1-4C)alkyl group and a (1-4C)alkoxy group.

“Pharmaceutically acceptable carrier” refers to a diluent, adjuvant,excipient or vehicle with, or in which a compound is administered.

“Treating” or “treatment” of any condition, such as an autoimmune,metabolic, allergic, cancer or infectious disease, refers, in certainembodiments, to ameliorating the condition (i.e., arresting or reducingthe development of the condition). In certain embodiments “treating” or“treatment” refers to ameliorating at least one physical parameter,which may not be discernible by the patient. In certain embodiments,“treating” or “treatment” refers to inhibiting the condition, eitherphysically. (e.g., stabilization of a discernible symptom),physiologically, (e.g., stabilization of a physical parameter), or both.In certain embodiments, “treating” or “treatment” refers to delaying theonset of the condition.

“Therapeutically effective amount” means the amount of a compound that,when administered to a patient for preventing or treating a conditionsuch as an autoimmune, metabolic, allergic, cancer or infectiousdisease, is sufficient to effect such treatment. The “therapeuticallyeffective amount” will vary depending on the compound, the condition andits severity and the age, weight, etc., of the patient.

DETAILED DESCRIPTION

Compounds for activating invariant natural killer T cells (iNKT) cellsare provided. Compounds according to certain embodiments activate iNKTcells and induce an increase in the production of one or more cytokines,such as IFNγ, IL-2, IL-4, IL-6 and TNFα. In some embodiments, activatediNKT cells are used to selectively reduce the presence of or eliminateinflammatory senescent cells, such as senescent cells having aninflammatory secretome (SASP). Methods for activating iNKT cells bycontacting an iNKT cell with an amount of the subject compounds andselectively reducing the presence of or eliminating senescent cells withactivated iNKT cells are also described. Compositions for practicing thesubject methods are also described.

Before the present invention is described in greater detail, it is to beunderstood that this invention is not limited to particular embodimentsdescribed, as such may, of course, vary. It is also to be understoodthat the terminology used herein is for the purpose of describingparticular embodiments only, and is not intended to be limiting, sincethe scope of the present invention will be limited only by the appendedclaims.

Where a range of values is provided, it is understood that eachintervening value, to the tenth of the unit of the lower limit unlessthe context clearly dictates otherwise, between the upper and lowerlimit of that range and any other stated or intervening value in thatstated range, is encompassed within the invention. The upper and lowerlimits of these smaller ranges may independently be included in thesmaller ranges and are also encompassed within the invention, subject toany specifically excluded limit in the stated range. Where the statedrange includes one or both of the limits, ranges excluding either orboth of those included limits are also included in the invention.

Certain ranges are presented herein with numerical values being precededby the term “about.” The term “about” is used herein to provide literalsupport for the exact number that it precedes, as well as a number thatis near to or approximately the number that the term precedes. Indetermining whether a number is near to or approximately a specificallyrecited number, the near or approximating unrecited number may be anumber which, in the context in which it is presented, provides thesubstantial equivalent of the specifically recited number.

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which this invention belongs. Although any methods andmaterials similar or equivalent to those described herein can also beused in the practice or testing of the present invention, representativeillustrative methods and materials are now described.

All publications and patents cited in this specification are hereinincorporated by reference as if each individual publication or patentwere specifically and individually indicated to be incorporated byreference and are incorporated herein by reference to disclose anddescribe the methods and/or materials in connection with which thepublications are cited. The citation of any publication is for itsdisclosure prior to the filing date and should not be construed as anadmission that the present invention is not entitled to antedate suchpublication by virtue of prior invention. Further, the dates ofpublication provided may be different from the actual publication dateswhich may need to be independently confirmed.

It is noted that, as used herein and in the appended claims, thesingular forms “a”, “an”, and “the” include plural referents unless thecontext clearly dictates otherwise. It is further noted that the claimsmay be drafted to exclude any optional element. As such, this statementis intended to serve as antecedent basis for use of such exclusiveterminology as “solely.” “only” and the like in connection with therecitation of claim elements, or use of a “negative” limitation.

As will be apparent to those of skill in the art upon reading thisdisclosure, each of the individual embodiments described and illustratedherein has discrete components and features which may be readilyseparated from or combined with the features of any of the other severalembodiments without departing from the scope or spirit of the presentinvention. Any recited method can be carried out in the order of eventsrecited or in any other order which is logically possible.

While the compounds and methods have or will be described for the sakeof grammatical fluidity with functional explanations, it is to beexpressly understood that the claims, unless expressly formulated under35 U.S.C. § 112, are not to be construed as necessarily limited in anyway by the construction of “means” or “steps” limitations, but are to beaccorded the full scope of the meaning and equivalents of the definitionprovided by the claims under the judicial doctrine of equivalents, andin the case where the claims are expressly formulated under 35 U.S.C. §112 are to be accorded full statutory equivalents under 35 U.S.C. § 112.

The publications discussed herein are provided solely for theirdisclosure prior to the filing date of the present application. Nothingherein is to be construed as an admission that the present invention isnot entitled to antedate such publication by virtue of prior invention.Further, the dates of publication provided may be different from theactual publication dates which may need to be independently confirmed.

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which this invention belongs. Although any methods andmaterials similar or equivalent to those described herein can also beused in the practice or testing of the present invention, the preferredmethods and materials are now described. All publications mentionedherein are incorporated herein by reference to disclose and describe themethods and/or materials in connection with which the publications arecited.

It is appreciated that certain features of the invention, which are, forclarity, described in the context of separate embodiments, may also beprovided in combination in a single embodiment. Conversely, variousfeatures of the invention, which are, for brevity, described in thecontext of a single embodiment, may also be provided separately or inany suitable sub-combination. All combinations of the embodimentspertaining to the chemical groups represented by the variables arespecifically embraced by the present invention and are disclosed hereinjust as if each and every combination was individually and explicitlydisclosed, to the extent that such combinations embrace compounds thatare stable compounds (i.e., compounds that can be isolated,characterised, and tested for biological activity). In addition, allsub-combinations of the chemical groups listed m the embodimentsdescribing such variables are also specifically embraced by the presentinvention and are disclosed herein just as if each and every suchsub-combination of chemical groups was individually and explicitlydisclosed herein.

Reference will now be made in detail to various embodiments. It will beunderstood that the invention is not limited to these embodiments. Tothe contrary, it is intended to cover alternatives, modifications, andequivalents as may be included within the spirit and scope of theallowed claims.

Compounds for Activating iNKT Cells and Selectively EliminatingSenescent Cells

Formula DCD-(I)

Aspects of the present disclosure according to certain embodimentsinclude a compound of formula DCD-(I):

-   -   wherein:    -   Z is selected from:

-   -   wherein        indicates the Z—O bond;    -   wherein X is —NHCO— or oxygen    -   R¹, R², R³ and R⁴ are each independently selected from hydrogen,        alkyl, substituted alkyl, heteroalkyl, substituted heteroalkyl,        cycloalkyl, substituted cycloalkyl, heterocycloalkyl,        substituted heterocycloalkyl, aryl, substituted aryl, arylalkyl,        substituted arylalkyl, heteroaryl, substituted heteroaryl,        heteroarylalkyl, and substituted heteroarylalkyl;    -   X₁ and X₂ are each independently selected from —C, —NR^(j), —O,        —SR^(k), —Si, wherein R^(j) and R^(k) are each independently        selected from hydrogen, alkyl or substituted alkyl, heteroalkyl,        substituted heteroalkyl, cycloalkyl, substituted cycloalkyl,        heterocycloalkyl, substituted heterocycloalkyl, aryl,        substituted aryl, arylalkyl, substituted arylalkyl, heteroaryl,        substituted heteroaryl, heteroarylalkyl, and substituted        heteroarylalkyl;    -   R^(a) is selected from hydrogen, oxygen, fluorine, —CF₃, or        together with X₂ form a cycloalkyl, substituted cycloalkyl,        heterocycloalkyl, substituted heterocycloalkyl, aryl,        substituted aryl, arylalkyl, substituted arylalkyl, heteroaryl,        substituted heteroaryl, heteroarylalkyl, and substituted        heteroarylalkyl;    -   wherein        indicates a double or single bond;    -   n is an integer from 2 to 25;    -   Y is selected from carbon, nitrogen or silicon;    -   R^(b), R^(c) and R^(d) are independently selected hydrogen,        alkyl, substituted alkyl, heteroalkyl, substituted heteroalkyl,        cycloalkyl, substituted cycloalkyl, heterocycloalkyl,        substituted heterocycloalkyl, aryl, substituted aryl, arylalkyl,        substituted arylalkyl, heteroaryl, substituted heteroaryl,        heteroarylalkyl, and substituted heteroarylalkyl, wherein when Y        is nitrogen, R^(d) is not present, or wherein R^(c) and R^(d)        together with Y form a cycloalkyl, substituted cycloalkyl, aryl,        substituted aryl, heteroaryl or substituted heteroaryl group;        and    -   R^(e) is alkyl or substituted alkyl,    -   or salt, solvate or hydrate thereof.

In embodiments, “salts” of the compounds of the present disclosure mayinclude: (1) acid addition salts, formed with inorganic acids such ashydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid,phosphoric acid, and the like; or formed with organic acids such asacetic acid, propionic acid, hexanoic acid, cyclopentanepropionic acid,glycolic acid, pyruvic acid, lactic acid, malonic acid, succinic acid,malic acid, maleic acid, fumaric acid, tartaric acid, citric acid,benzoic acid, 3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, mandelicacid, methanesulfonic acid, ethanesulfonic acid, 1,2-ethane-disulfonicacid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid,4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid,4-toluenesulfonic acid, camphorsulfonic acid,4-methylbicyclo[2.2.2]-oct-2-ene-1-carboxylic acid, glucoheptonic acid,3-phenylpropionic acid, trimethylacetic acid, tertiary butylacetic acid,lauryl sulfuric acid, gluconic acid, glutamic acid, hydroxynaphthoicacid, salicylic acid, stearic acid, muconic acid, and the like; or (2)salts formed when an acidic proton present in the compound is replacedby a metal ion, e.g., an alkali metal ion, an alkaline earth ion, or analuminum ion; or coordinates with an organic base such as ethanolamine,diethanolamine, triethanolamine, N-methylglucamine and the like.

The term “solvate” as used herein refers to a complex or aggregateformed by one or more molecules of a solute, e.g. a compound of DCD-(I)or a salt thereof, and one or more molecules of a solvent. Such solvatesmay be crystalline solids having a substantially fixed molar ratio ofsolute and solvent. Representative solvents include by way of example,water, methanol, ethanol, isopropanol, acetic acid, and the like. Whenthe solvent is water, the solvate formed is a hydrate.

Formula DCD-(IA-1)

Aspects of the present disclosure according to certain embodimentsinclude a compound of formula DCD-(IA-1):

-   -   wherein:    -   wherein X is —NHCO— or oxygen    -   R¹, R², R³ and R⁴ are each independently selected from hydrogen,        alkyl, substituted alkyl, heteroalkyl, substituted heteroalkyl,        cycloalkyl, substituted cycloalkyl, heterocycloalkyl,        substituted heterocycloalkyl, aryl, substituted aryl, arylalkyl,        substituted arylalkyl, heteroaryl, substituted heteroaryl,        heteroarylalkyl, and substituted heteroarylalkyl;    -   X₁ and X₂ are each independently selected from —C, —NR^(j), —O,        —SR^(k), —Si, wherein R^(j) and R^(k) are each independently        selected from hydrogen, alkyl or substituted alkyl, heteroalkyl,        substituted heteroalkyl, cycloalkyl, substituted cycloalkyl,        heterocycloalkyl, substituted heterocycloalkyl, aryl,        substituted aryl, arylalkyl, substituted arylalkyl, heteroaryl,        substituted heteroaryl, heteroarylalkyl, and substituted        heteroarylalkyl;    -   R^(a) is selected from hydrogen, oxy gen, fluorine, —CF₃, or        together with X₂ form a cycloalkyl, substituted cycloalkyl,        heterocycloalkyl, substituted heterocycloalkyl, aryl,        substituted aryl, arylalkyl, substituted arylalkyl, heteroaryl,        substituted heteroaryl, heteroarylalkyl, and substituted        heteroarylalkyl;    -   wherein        indicates a double or single bond;    -   n is an integer from 2 to 25;    -   Y is selected from carbon, nitrogen or silicon;    -   R^(b), R^(c) and R^(d) are independently selected hydrogen,        alkyl, substituted alkyl, heteroalkyl, substituted heteroalkyl,        cycloalkyl, substituted cycloalkyl, heterocycloalkyl,        substituted heterocycloalkyl, aryl, substituted aryl, arylalkyl,        substituted arylalkyl, heteroaryl, substituted heteroaryl,        heteroarylalkyl, and substituted heteroarylalkyl, wherein when Y        is nitrogen, R^(d) is not present, or wherein R^(c) and R^(d)        together with Y form a cycloalkyl, substituted cycloalkyl, aryl,        substituted aryl, heteroaryl or substituted heteroaryl group;        and    -   R^(e) is alkyl or substituted alkyl,    -   or salt, solvate or hydrate thereof.

In some instances, R¹ is hydrogen. In some instances, R² is hydrogen. Insome instances, R³ is hydrogen. In some instances, R⁴ is hydrogen. Incertain instances, each of R¹, R², R³ and R⁴ are hydrogen.

In other instances, R¹ is:

-   -   wherein        represents the R¹—O bond;    -   R⁸ is hydrogen, alkyl or substituted alkyl; In some instances,        R⁸ is selected from hydrogen, methyl, ethyl and cyclopropyl.    -   R⁹ is —NR^(f) or —OR^(f) wherein R^(f) is alkyl, substituted        alkyl, acyl, alkylacyl or substituted alkylacyl, or wherein        R^(f) together with R¹⁰ form a cycloalkyl, substituted        cycloalkyl, heterocycloalkyl, substituted heterocycloalkyl,        aryl, substituted aryl, arylalkyl, substituted arylalkyl,        heteroaryl, substituted heteroaryl, heteroarylalkyl, and        substituted heteroarylalkyl; and    -   R¹⁰ is cycloalkyl, substituted cycloalkyl, heterocycloalkyl,        substituted heterocycloalkyl, aryl, substituted aryl, arylalkyl,        substituted arylalkyl, heteroaryl, substituted heteroaryl,        heteroarylalkyl, and substituted heteroarylalkyl or wherein R¹⁰        together with R^(f) form a cycloalkyl, substituted cycloalkyl,        heterocycloalkyl, substituted heterocycloalkyl, aryl,        substituted aryl, arylalkyl, substituted arylalkyl, heteroaryl,        substituted heteroaryl, heteroarylalkyl, and substituted        heteroarylalkyl.

In certain instances, R¹ is selected from:

-   -   wherein        represents the R¹—O bond;    -   R⁸ is hydrogen, alkyl, substituted alkyl; and    -   R^(g) is hydrogen or a halogen selected from fluorine, chlorine,        bromine or iodine.

In other instances, R¹ is:

-   -   wherein:    -   R¹¹ is alkyl or substituted alkyl;    -   R¹² is alkyl or alkyl substituted with a cycloalkyl, substituted        cycloalkyl, heterocycloalkyl, substituted heterocycloalkyl,        aryl, substituted aryl, arylalkyl, substituted arylalkyl,        heteroaryl, substituted heteroaryl, heteroarylalkyl, and        substituted heteroarylalkyl.

In some instances, R¹¹ is selected from methyl, ethyl, n-propyl,iso-propyl, n-butyl, iso-butyl, t-butyl. In certain instances, R¹¹ isiso-propyl.

In certain instances, R¹ is:

-   -   wherein R^(h) and R^(i) are each independently selected from        hydrogen, hydroxyl or halogen. In some instances, R^(h) is        hydroxyl. In some instances, R^(h) is F. In some instances,        R^(h) is Cl. In some instances, R^(h) is I. In some instances,        R^(h) is Br. In some instances, R^(i) is hydroxyl. In some        instances, R^(i) is F. In other instances, R^(i) is Cl. In other        instances, R^(i) is I. In other instances, R^(i) is Br.

Formula DCD-(IA-2)

Aspects of the present disclosure according to certain embodimentsinclude a compound of formula DCD-(IA-2):

-   -   wherein:    -   wherein X is —NHCO— or oxygen;    -   R¹, R² and R³ are each independently selected from hydrogen,        alkyl, substituted alkyl, heteroalkyl, substituted heteroalkyl,        cycloalkyl, substituted cycloalkyl, heterocycloalkyl,        substituted heterocycloalkyl, aryl, substituted aryl, arylalkyl,        substituted arylalkyl, heteroaryl, substituted heteroaryl,        heteroarylalkyl, and substituted heteroarylalkyl;    -   X₁ and X₂ are each independently selected from —C, —NR^(j), —O,        —SR^(k), —Si, wherein R^(j) and R^(k) are each independently        selected from hydrogen, alkyl or substituted alkyl, heteroalkyl,        substituted heteroalkyl, cycloalkyl, substituted cycloalkyl,        heterocycloalkyl, substituted heterocycloalkyl, aryl,        substituted aryl, arylalkyl, substituted arylalkyl, heteroaryl,        substituted heteroaryl, heteroarylalkyl, and substituted        heteroarylalkyl;    -   R^(a) is selected from hydrogen, oxygen, fluorine, —CF₃, or        together with X₂ form a cycloalkyl, substituted cycloalkyl,        heterocycloalkyl, substituted heterocycloalkyl, aryl,        substituted aryl, arylalkyl, substituted arylalkyl, heteroaryl,        substituted heteroaryl, heteroarylalkyl, and substituted        heteroarylalkyl;    -   wherein        indicates a double or single bond;    -   n is an integer from 2 to 25;    -   Y is selected from carbon, nitrogen or silicon;    -   R^(b), R^(c) and R^(d) are independently selected hydrogen,        alkyl, substituted alkyl, heteroalkyl, substituted heteroalkyl,        cycloalkyl, substituted cycloalkyl, heterocycloalkyl,        substituted heterocycloalkyl, aryl, substituted aryl, arylalkyl,        substituted arylalkyl, heteroaryl, substituted heteroaryl,        heteroarylalkyl, and substituted heteroarylalkyl, wherein when Y        is nitrogen, R^(d) is not present, or wherein R^(c) and R^(d)        together with Y form a cycloalkyl, substituted cycloalkyl, aryl,        substituted aryl, heteroaryl or substituted heteroaryl group;        and    -   R^(c) is alkyl or substituted alkyl,    -   or salt, solvate or hydrate thereof.

In some instances, R¹ is hydrogen. In some instances, R² is hydrogen. Insome instances. R³ is hydrogen. In certain instances, each of R¹, R² andR³ are hydrogen.

In other instances, R¹ is:

-   -   wherein        represents the R¹—O bond;    -   R⁸ is hydrogen, alkyl or substituted alkyl; In some instances,        R⁸ is selected from hydrogen, methyl, ethyl and cyclopropyl    -   R⁹ is —NR^(f) or —OR, wherein R^(f) is alkyl, substituted alkyl,        acyl, alkylacyl or substituted alkylacyl, or wherein R^(f)        together with R¹⁰ form a cycloalkyl, substituted cycloalkyl,        heterocycloalkyl, substituted heterocycloalkyl, aryl,        substituted aryl, arylalkyl, substituted arylalkyl, heteroaryl,        substituted heteroaryl, heteroarylalkyl, and substituted        heteroarylalkyl; and    -   R¹⁰ is cycloalkyl, substituted cycloalkyl, heterocycloalkyl,        substituted heterocycloalkyl, aryl, substituted aryl, arylalkyl,        substituted arylalkyl, heteroaryl, substituted heteroaryl,        heteroarylalkyl, and substituted heteroarylalkyl or wherein R¹⁰        together with R^(f) form a cycloalkyl, substituted cycloalkyl,        heterocycloalkyl, substituted heterocycloalkyl, aryl,        substituted aryl, arylalkyl, substituted arylalkyl, heteroaryl,        substituted heteroaryl, heteroarylalkyl, and substituted        heteroarylalkyl.

In certain instances, R¹ is selected from:

-   -   wherein        represents the R¹—O bond;    -   R^(g) is hydrogen, alkyl, substituted alkyl; and    -   R^(g) is hydrogen or a halogen selected from fluorine, chlorine,        bromine or iodine.    -   In other instances, R¹ is:

-   -   wherein:    -   R¹¹ is alkyl or substituted alkyl;    -   R¹² is alkyl or alkyl substituted with a cycloalkyl, substituted        cycloalkyl, heterocycloalkyl, substituted heterocycloalkyl,        aryl, substituted aryl, arylalkyl, substituted arylalkyl,        heteroaryl, substituted heteroaryl, heteroarylalkyl, and        substituted heteroarylalkyl.

In some instances, R¹¹ is selected from methyl, ethyl, n-propyl,iso-propyl, n-butyl, iso-butyl, t-butyl. In certain instances, R¹¹ isiso-propyl.

In certain instances, R¹ is:

-   -   wherein R^(h) and R^(i) are each independently selected from        hydrogen, hydroxyl or halogen. In some instances, R^(h) is        hydroxyl. In some instances, R^(h) is F. In some instances,        R^(h) is Cl. In some instances, R^(h) is I. In some instances,        R^(h) is Br. In some instances, R^(i) is hydroxyl. In some        instances, R^(i) is F. In other instances, R^(i) is Cl. In other        instances, R^(i) is I. In other instances, R^(i) is Br.

Formula DCD-(IA-3)

Aspects of the present disclosure according to certain embodimentsinclude a compound of formula DCD-(IA-3):

-   -   wherein:    -   wherein X is —NHCO— or oxygen;    -   R¹, R², R³ and R⁴ are each independently selected from hydrogen,        alkyl, substituted alkyl, heteroalkyl, substituted heteroalkyl,        cycloalkyl, substituted cycloalkyl, heterocycloalkyl,        substituted heterocycloalkyl, aryl, substituted aryl, arylalkyl,        substituted arylalkyl, heteroaryl, substituted heteroaryl,        heteroarylalkyl, and substituted heteroarylalkyl;    -   X₁ and X₂ are each independently selected from —C, —NR^(j), —O,        —SR^(k), —Si, wherein R^(j) and R^(k) are each independently        selected from hydrogen, alkyl or substituted alkyl, heteroalkyl,        substituted heteroalkyl, cycloalkyl, substituted cycloalkyl,        heterocycloalkyl, substituted heterocycloalkyl, aryl,        substituted aryl, arylalkyl, substituted arylalkyl, heteroaryl,        substituted heteroaryl, heteroarylalkyl, and substituted        heteroarylalkyl;    -   R^(a) is selected from hydrogen, oxygen, fluorine, —CF₃, or        together with X₂ form a cycloalkyl, substituted cycloalkyl,        heterocycloalkyl, substituted heterocycloalkyl, aryl,        substituted aryl, arylalkyl, substituted arylalkyl, heteroaryl,        substituted heteroaryl, heteroarylalkyl, and substituted        heteroarylalkyl;    -   wherein        indicates a double or single bond;    -   n is an integer from 2 to 25,    -   Y is selected from carbon, nitrogen or silicon;    -   R^(b), R^(c) and R^(d) are independently selected hydrogen,        alkyl, substituted alkyl, heteroalkyl, substituted heteroalkyl,        cycloalkyl, substituted cycloalkyl, heterocycloalkyl,        substituted heterocycloalkyl, aryl, substituted aryl, arylalkyl,        substituted arylalkyl, heteroaryl, substituted heteroaryl,        heteroarylalkyl, and substituted heteroarylalkyl, wherein when Y        is nitrogen, R^(d) is not present, or wherein R^(c) and R^(d)        together with Y form a cycloalkyl, substituted cycloalkyl, aryl,        substituted aryl, heteroaryl or substituted heteroaryl group;        and    -   R^(e) is alkyl or substituted alkyl,    -   or salt, solvate or hydrate thereof.

In some instances, R¹ is hydrogen. In some instances, R² is hydrogen. Insome instances, R³ is hydrogen. In some instances, R⁴ is hydrogen. Incertain instances, each of R¹, R², R³ and R⁴ are hydrogen.

In other instances, R¹ is:

-   -   wherein        represents the R¹—O bond;    -   R⁸ is hydrogen, alkyl or substituted alkyl; In some instances,        R⁸ is selected from hydrogen, methyl, ethyl and cyclopropyl.    -   R⁹ is —NR^(f) or —OR^(f), wherein R^(f) is alkyl, substituted        alkyl, acyl, alkylacyl or substituted alkylacyl, or wherein        R^(f) together with R¹⁰ form a cycloalkyl, substituted        cycloalkyl, heterocycloalkyl, substituted heterocycloalkyl,        aryl, substituted aryl, arylalkyl, substituted arylalkyl,        heteroaryl, substituted heteroaryl, heteroarylalkyl, and        substituted heteroarylalkyl; and    -   R¹⁰ is cycloalkyl, substituted cycloalkyl, heterocycloalkyl,        substituted heterocycloalkyl, aryl, substituted aryl, arylalkyl,        substituted arylalkyl, heteroaryl, substituted heteroaryl,        heteroarylalkyl, and substituted heteroarylalkyl or wherein R¹⁰        together with R^(f) form a cycloalkyl, substituted cycloalkyl,        heterocycloalkyl, substituted heterocycloalkyl, aryl,        substituted aryl, arylalkyl, substituted arylalkyl, heteroaryl,        substituted heteroaryl, heteroarylalkyl, and substituted        heteroarylalkyl.

In certain instances, R¹ is selected from:

-   -   wherein        represents the R¹—O bond;    -   R⁸ is hydrogen, alkyl, substituted alkyl; and    -   R^(g) is hydrogen or a halogen selected from fluorine, chlorine,        bromine or iodine.

In other instances, R¹ is:

-   -   wherein:    -   R¹¹ is alkyl or substituted alkyl;    -   R¹² is alkyl or alkyl substituted with a cycloalkyl, substituted        cycloalkyl, heterocycloalkyl, substituted heterocycloalkyl,        aryl, substituted aryl, arylalkyl, substituted arylalkyl,        heteroaryl, substituted heteroaryl, heteroarylalkyl, and        substituted heteroarylalkyl.

In some instances, R¹¹ is selected from methyl, ethyl, n-propyl,iso-propyl, n-butyl, iso-butyl, t-butyl. In certain instances, R¹¹ isiso-propyl.

In certain instances, R¹ is:

-   -   wherein R^(h) and R^(i) are each independently selected from        hydrogen, hydroxyl or halogen. In some instances, R^(h) is        hydroxyl. In some instances, R^(h) is F. In some instances,        R^(h) is Cl. In some instances, R^(h) is 1. In some instances,        R^(h) is Br. In some instances, R^(i) is hydroxyl. In some        instances, R^(h) is F. In other instances, R^(h) is Cl. In other        instances, R^(i) is I.

In other instances, R^(i) is Br.

Formula DCD-(IA-4)

Aspects of the present disclosure according to certain embodimentsinclude a compound of formula DCD-(IA-4):

-   -   wherein:    -   wherein X is —NHCO— or oxygen;    -   R¹ and R² are each independently selected from hydrogen, alkyl,        substituted alkyl, heteroalkyl, substituted heteroalkyl,        cycloalkyl, substituted cycloalkyl, heterocycloalkyl,        substituted heterocycloalkyl, aryl, substituted aryl, arylalkyl,        substituted arylalkyl, heteroaryl, substituted heteroaryl,        heteroarylalkyl, and substituted heteroarylalkyl;    -   X₁ and X₂ are each independently selected from —C, —NR^(j), —O,        —SR^(k), —Si, wherein R^(j) and R^(k) are each independently        selected from hydrogen, alkyl or substituted alkyl, heteroalkyl,        substituted heteroalkyl, cycloalkyl, substituted cycloalkyl,        heterocycloalkyl, substituted heterocycloalkyl, aryl,        substituted aryl, arylalkyl, substituted arylalkyl, heteroaryl,        substituted heteroaryl, heteroarylalkyl, and substituted        heteroarylalkyl;    -   R^(a) is selected from hydrogen, oxygen, fluorine, —CF₃, or        together with X₂ form a cycloalkyl, substituted cycloalkyl,        heterocycloalkyl, substituted heterocycloalkyl, aryl,        substituted aryl, arylalkyl, substituted arylalkyl, heteroaryl,        substituted heteroaryl, heteroarylalkyl, and substituted        heteroarylalkyl;    -   wherein        indicates a double or single bond;    -   n is an integer from 2 to 25;    -   Y is selected from carbon, nitrogen or silicon;    -   R^(b), R^(c) and R^(d) are independently selected hydrogen,        alkyl, substituted alkyl, heteroalkyl, substituted heteroalkyl,        cycloalkyl, substituted cycloalkyl, heterocycloalkyl,        substituted heterocycloalkyl, aryl, substituted aryl, arylalkyl,        substituted arylalkyl, heteroaryl, substituted heteroaryl,        heteroarylalkyl, and substituted heteroarylalkyl, wherein when Y        is nitrogen, R^(d) is not present, or wherein R^(c) and R^(d)        together with Y form a cycloalkyl, substituted cycloalkyl, aryl,        substituted aryl, heteroaryl or substituted heteroaryl group;        and    -   R^(e) is alkyl or substituted alkyl, or salt, solvate or hydrate        thereof.

In some instances, R¹ is hydrogen. In some instances, R² is hydrogen. Insome instances, each of R¹ and R² are each hydrogen.

In other instances, R¹ is:

-   -   wherein        a represents the R¹—O bond;    -   R⁸ is hydrogen, alkyl or substituted alkyl; In some instances,        R⁸ is selected from hydrogen, methyl, ethyl and cyclopropyl.    -   R⁹ is —NR^(f) or —OR^(f), wherein R^(f) is alkyl, substituted        alkyl, acyl, alkylacyl or substituted alkylacyl, or wherein R¹        together with R¹⁰ form a cycloalkyl, substituted cycloalkyl,        heterocycloalkyl, substituted heterocycloalkyl, aryl,        substituted aryl, arylalkyl, substituted arylalkyl, heteroaryl,        substituted heteroaryl, heteroarylalkyl, and substituted        heteroarylalkyl; and    -   R¹⁰ is cycloalkyl, substituted cycloalkyl, heterocycloalkyl,        substituted heterocycloalkyl, aryl, substituted aryl, arylalkyl,        substituted arylalkyl, heteroaryl, substituted heteroaryl,        heteroarylalkyl, and substituted heteroarylalkyl or wherein R¹⁰        together with R^(f) form a cycloalkyl, substituted cycloalkyl,        heterocycloalkyl, substituted heterocycloalkyl, aryl,        substituted aryl, arylalkyl, substituted arylalkyl, heteroaryl,        substituted heteroaryl, heteroarylalkyl, and substituted        heteroarylalkyl.

In certain instances, R¹ is selected from:

-   -   wherein        represents the R¹—O bond;    -   R⁸ is hydrogen, alkyl, substituted alkyl; and    -   R^(g) is hydrogen or a halogen selected from fluorine, chlorine,        bromine or iodine.

In other instances, R¹ is:

-   -   wherein:    -   R¹¹ is alkyl or substituted alkyl;    -   R¹² is alkyl or alkyl substituted with a cycloalkyl, substituted        cycloalkyl, heterocycloalkyl, substituted heterocycloalkyl,        aryl, substituted aryl, arylalkyl, substituted arylalkyl,        heteroaryl, substituted heteroaryl, heteroarylalkyl, and        substituted heteroarylalkyl.

In some instances, R¹¹ is selected from methyl, ethyl, n-propyl,iso-propyl, n-butyl, iso-butyl, t-butyl. In certain instances, R¹¹ isiso-propyl.

In certain instances, R¹ is:

-   -   wherein R^(h) and R^(i) are each independently selected from        hydrogen, hydroxyl or halogen. In some instances, R^(h) is        hydroxyl. In some instances, R^(h) is F. In some instances,        R^(h) is Cl. In some instances, R^(h) is I. In some instances,        R^(h) is Br. In some instances, R^(i) is hydroxyl. In some        instances, R^(i) is F. In other instances, R^(i) is Cl. In other        instances, R^(i) is I. In other instances, R^(i) is Br.

Formula DCD-(IB-1)

Aspects of the present disclosure according to certain embodimentsinclude a compound of formula DCD-(IB-1):

-   -   wherein:    -   Z is selected from:

-   -   wherein        indicates the Z—O bond;    -   wherein X is —NHCO— or oxygen;    -   R¹, R², R³ and R⁴ are each independently selected from hydrogen,        alkyl, substituted alkyl, heteroalkyl, substituted heteroalkyl,        cycloalkyl, substituted cycloalkyl, heterocycloalkyl,        substituted heterocycloalkyl, aryl, substituted aryl, arylalkyl,        substituted arylalkyl, heteroaryl, substituted heteroaryl,        heteroarylalkyl, and substituted heteroarylalkyl;    -   n is an integer from 2 to 25;    -   Y is selected from carbon, nitrogen or silicon    -   R^(b), R^(c) and R^(d) are independently selected hydrogen,        alkyl, substituted alkyl, heteroalkyl, substituted heteroalkyl,        cycloalkyl, substituted cycloalkyl, heterocycloalkyl,        substituted heterocycloalkyl, aryl, substituted aryl, arylalkyl,        substituted arylalkyl, heteroaryl, substituted heteroaryl,        heteroarylalkyl, and substituted heteroarylalkyl, wherein when Y        is nitrogen, R^(d) is not present, or wherein R^(c) and R^(d)        together with Y form a cycloalkyl, substituted cycloalkyl, aryl,        substituted aryl, heteroaryl or substituted heteroaryl group;        and    -   R^(e) is alkyl or substituted alkyl,    -   or salt, solvate or hydrate thereof.

Formula DCD-(IB-2)

Aspects of the present disclosure according to certain embodimentsinclude a compound of formula DCD-(IB-2):

-   -   wherein:    -   Z is selected from:

-   -   wherein        indicates the Z—O bond;    -   wherein X is —NHCO— or oxygen;    -   R¹, R², R³ and R⁴ are each independently selected from hydrogen,        alkyl, substituted alkyl, heteroalkyl, substituted heteroalkyl,        cycloalkyl, substituted cycloalkyl, heterocycloalkyl,        substituted heterocycloalkyl, aryl, substituted aryl, arylalkyl,        substituted arylalkyl, heteroaryl, substituted heteroaryl,        heteroarylalkyl, and substituted heteroarylalkyl;    -   R^(k) is selected from hydrogen, alkyl, substituted alkyl,        heteroalkyl, substituted heteroalkyl, cycloalkyl, substituted        cycloalkyl, heterocycloalkyl, substituted heterocycloalkyl,        aryl, substituted aryl, arylalkyl, substituted arylalkyl,        heteroaryl, substituted heteroaryl, heteroarylalkyl, and        substituted heteroarylalkyl. In some instances, R^(k) is        selected from methyl, ethyl, n-propyl, iso-propyl, n-butyl,        iso-butyl, t-butyl. In certain instances, R^(k) is methyl.    -   N is an integer from 2 to 25;    -   Y is selected from carbon, nitrogen or silicon;    -   R^(b), R^(c) and R^(d) are independently selected hydrogen,        alkyl, substituted alkyl, heteroalkyl, substituted heteroalkyl,        cycloalkyl, substituted cycloalkyl, heterocycloalkyl,        substituted heterocycloalkyl, aryl, substituted aryl, arylalkyl,        substituted arylalkyl, heteroaryl, substituted heteroaryl,        heteroarylalkyl, and substituted heteroarylalkyl, wherein when Y        is nitrogen, R^(d) is not present, or wherein R^(c) and R^(d)        together with Y form a cycloalkyl, substituted cycloalkyl, aryl,        substituted aryl, heteroaryl or substituted heteroaryl group;        and    -   R^(e) is alkyl or substituted alkyl,    -   or salt, solvate or hydrate thereof.

Formula DCD-(IB-3)

Aspects of the present disclosure according to certain embodimentsinclude a compound of formula DCD-(IB-3):

-   -   wherein:    -   Z is selected from:

-   -   wherein        indicates the Z—O bond;    -   wherein X is —NHCO— or oxygen;    -   R¹, R², R³ and R⁴ are each independently selected from hydrogen,        alkyl, substituted alkyl, heteroalkyl, substituted heteroalkyl,        cycloalkyl, substituted cycloalkyl, heterocycloalkyl,        substituted heterocycloalkyl, aryl, substituted aryl, arylalkyl,        substituted arylalkyl, heteroaryl, substituted heteroaryl,        heteroarylalkyl, and substituted heteroarylalkyl;    -   n is an integer from 2 to 25;    -   Y is selected from carbon, nitrogen or silicon;    -   R^(b), R^(c) and R^(d) are independently selected hydrogen,        alkyl, substituted alkyl, heteroalkyl, substituted heteroalkyl,        cycloalkyl, substituted cycloalkyl, heterocycloalkyl,        substituted heterocycloalkyl, aryl, substituted aryl, arylalkyl,        substituted arylalkyl, heteroaryl, substituted heteroaryl,        heteroarylalkyl, and substituted heteroarylalkyl, wherein when Y        is nitrogen, R^(d) is not present, or wherein R^(c) and R^(d)        together with Y form a cycloalkyl, substituted cycloalkyl, aryl,        substituted aryl, heteroaryl or substituted heteroaryl group;        and    -   R^(e) is alkyl or substituted alkyl,    -   or salt, solvate or hydrate thereof.

Formula DCD-(IB-4)

Aspects of the present disclosure according to certain embodimentsinclude a compound of formula DCD-(IB-4):

-   -   wherein:    -   Z is selected from:

-   -   wherein        indicates Z—O bond;    -   wherein X is —NHCO— or oxygen;    -   R¹, R², R³ and R⁴ are each independently selected from hydrogen,        alkyl, substituted alkyl, heteroalkyl, substituted heteroalkyl,        cycloalkyl, substituted cycloalkyl, heterocycloalkyl,        substituted heterocycloalkyl, aryl, substituted aryl, arylalkyl,        substituted arylalkyl, heteroaryl, substituted heteroaryl,        heteroarylalkyl, and substituted heteroarylalkyl;    -   n is an integer from 2 to 25;    -   Y is selected from carbon, nitrogen or silicon;    -   R^(b), R^(c) and R^(d) are independently selected hydrogen,        alkyl, substituted alkyl, heteroalkyl, substituted heteroalkyl,        cycloalkyl, substituted cycloalkyl, heterocycloalkyl,        substituted heterocycloalkyl, aryl, substituted aryl, arylalkyl,        substituted arylalkyl, heteroaryl, substituted heteroaryl,        heteroarylalkyl, and substituted heteroarylalkyl, wherein when Y        is nitrogen, R^(d) is not present, or wherein R^(c) and R^(d)        together with Y form a cycloalkyl, substituted cycloalkyl, aryl,        substituted aryl, heteroaryl or substituted heteroaryl group;        and    -   R^(e) is alkyl or substituted alkyl,    -   or salt, solvate or hydrate thereof.

Formula DCD-(IX-5)

Aspects of the present disclosure according to certain embodimentsinclude a compound of formula DCD-(IB-5):

-   -   wherein:    -   Z is selected from:

-   -   wherein        indicates the Z—O bond;    -   wherein X is —NHCO— or oxygen;    -   R¹, R², R³ and R⁴ are each independently selected from hydrogen,        alkyl, substituted alkyl, heteroalkyl, substituted heteroalkyl,        cycloalkyl, substituted cycloalkyl, heterocycloalkyl,        substituted heterocycloalkyl, aryl, substituted aryl, arylalkyl,        substituted arylalkyl, heteroaryl, substituted heteroaryl,        heteroarylalkyl, and substituted heteroarylalkyl;    -   n is an integer from 2 to 25;    -   Y is selected from carbon, nitrogen or silicon;    -   R^(b), R^(c) and R^(d) are independently selected hydrogen,        alkyl, substituted alkyl, heteroalkyl, substituted heteroalkyl,        cycloalkyl, substituted cycloalkyl, heterocycloalkyl,        substituted heterocycloalkyl, aryl, substituted aryl, arylalkyl,        substituted arylalkyl, heteroaryl, substituted heteroaryl,        heteroarylalkyl, and substituted heteroarylalkyl, wherein when Y        is nitrogen, R^(d) is not present, or wherein R^(c) and R^(d)        together with Y form a cycloalkyl, substituted cycloalkyl, aryl,        substituted aryl, heteroaryl or substituted heteroaryl group;        and    -   R^(e) is alkyl or substituted alkyl,    -   or salt, solvate or hydrate thereof.

In embodiments of compounds of formulae DCD-(I), DCD-(IA) and DCD-(IB),in some instances, R^(e) may be alkyl or substituted alkyl. In someinstances, R^(e) is a C8 to C20 alkyl. In some instances, R^(e) is asubstituted C8 to C20 alkyl. In certain instances, R^(e) is a C13 alkyl.

In some instances n is an integer from 2 to 25, such as where n isselected from 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17,18, 19, 20, 21, 22, 23, 24 and 25.

In some instances, Y is carbon. In some instances, Y is nitrogen. Insome instances, Y is silicon.

In some instances, R^(b) is hydrogen. In some instances, R^(b) is alkyl.In some instances, R^(b) is a C₁-C₁₆ alkyl. In some instances, R^(b) isselected from methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl,tert-butyl, pentyl, hexyl, heptyl and octyl. In some instances, R^(b) isselected from:

Wherein

indicates a bond to Y and R^(z) is hydrogen, alkyl or alkyl substitutedwith a cycloalkyl, substituted cycloalkyl, heterocycloalkyl, substitutedheterocycloalkyl, aryl, substituted aryl, arylalkyl, substitutedarylalkyl, heteroaryl, substituted heteroaryl, heteroarylalkyl, andsubstituted heteroarylalkyl. In certain instances, R^(z) is alkyl, suchas a C1 to C16 alkyl or C1 to C16 substituted alkyl.

In some instances, R^(c) is hydrogen. In some instances, R^(c) is alkyl.In some instances, R^(c) is a C1-C16 alkyl. In some instances, R^(c) isselected from methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl,tert-butyl, pentyl, hexyl, heptyl and octyl.

In some embodiments, R^(d) is a C5 to C25 alkyl or a C5 to C25 alkylsubstituted with a cycloalkyl group, substituted cycloalkyl group,heterocycloalkyl group, substituted heterocycloalkyl group, aryl group,substituted aryl group, arylalkyl group, substituted arylalkyl group,heteroaryl group, substituted heteroaryl group, heteroarylalkyl group,or substituted heteroarylalkyl group. In some instances, R^(d) is a C5to C25 alkyl substituted with a moiety selected from the groupconsisting of

wherein

indicates the bond to the C5 to C25 alkyl; and R^(m) and R^(n) areindependently selected from hydrogen, halogen, hydroxyl, substitutedhydroxyl, amino, substituted amino, thiol, substituted thiol, sulfoxide,substituted sulfoxide, sulfone, substituted sulfone, sulfoximine,substituted sulfoximine, acyl, aminoacyl, alkyl, substituted alkyl,heteroalkyl, substituted heteroalkyl, cycloalkyl, substitutedcycloalkyl, spiroalkyl, heterocycloalkyl, substituted heterocycloalkyl,aryl, substituted aryl, arylalkyl, substituted arylalkyl, heteroaryl,substituted heteroaryl, heteroarylalkyl, and substitutedheteroarylalkyl.

In some instances, R^(m) is hydrogen. In some instances, R^(m) ishalogen. In some instances, R^(m) is selected from fluorine, bromine oriodine. In some instances, R^(n) is hydrogen. In some instances, R^(n)is halogen. In some instances, R^(n) is fluorine, bromine or iodine.

Formula DCD-(II)

Aspects of the present disclosure according to certain embodimentsinclude a compound of formulae DCD-(II):

-   -   wherein:    -   Z is selected from:

-   -   wherein        indicates the Z—O bond;    -   wherein X is —NHCO— or oxygen;    -   R¹, R², R³ and R⁴ are each independently selected from hydrogen,        alkyl, substituted alkyl, heteroalkyl, substituted heteroalkyl,        cycloalkyl, substituted cycloalkyl, heterocycloalkyl,        substituted heterocycloalkyl, aryl, substituted aryl, arylalkyl,        substituted arylalkyl, heteroaryl, substituted heteroaryl,        heteroarylalkyl, and substituted heteroarylalkyl;    -   X₁, X₂, X₃, X₄ and X₅ are each independently selected from        carbon, nitrogen, oxygen or sulfur;    -   R^(a) is optionally absent or when present is selected from        hydrogen or oxygen;    -   wherein        indicates a double or single bond,    -   n is an integer from 2 to 25;    -   Y is selected from carbon, nitrogen or silicon;    -   R^(b), R^(c) and R^(d) are independently selected hydrogen,        alkyl, substituted alkyl, heteroalkyl, substituted heteroalkyl,        cycloalkyl, substituted cycloalkyl, heterocycloalkyl,        substituted heterocycloalkyl, aryl, substituted aryl, arylalkyl,        substituted arylalkyl, heteroaryl, substituted heteroaryl,        heteroarylalkyl, and substituted heteroarylalkyl, wherein when Y        is nitrogen, R^(d) is not present, or wherein R^(c) and R^(d)        together with Y form a cycloalkyl, substituted cycloalkyl, aryl,        substituted aryl, heteroaryl or substituted heteroaryl group;        and    -   R^(e) is alkyl or substituted alkyl,    -   or salt, solvate or hydrate thereof.

Formula DCD-(IIA-1)

Aspects of the present disclosure according to certain embodimentsinclude a compound of formula DCD-(IIA-1):

-   -   wherein X is —NHCO— or oxygen;    -   R¹, R², R³ and R⁴ are each independently selected from hydrogen,        alkyl, substituted alkyl, heteroalkyl, substituted heteroalkyl,        cycloalkyl, substituted cycloalkyl, heterocycloalkyl,        substituted heterocycloalkyl, aryl, substituted aryl, arylalkyl,        substituted arylalkyl, heteroaryl, substituted heteroaryl,        heteroarylalkyl, and substituted heteroarylalkyl;    -   X₁, X₂, X₃, X₄ and X₅ are each independently selected from        carbon, nitrogen, oxygen or sulfur;    -   R^(a) is optionally absent or when present is selected from        hydrogen or oxygen;    -   wherein        indicates a double or single bond;    -   n is an integer from 2 to 25;    -   Y is selected from carbon, nitrogen or silicon;    -   R^(b), R^(c) and R^(d) are independently selected hydrogen,        alkyl, substituted alkyl, heteroalkyl, substituted heteroalkyl,        cycloalkyl, substituted cycloalkyl, heterocycloalkyl,        substituted heterocycloalkyl, aryl, substituted aryl, arylalkyl,        substituted arylalkyl, heteroaryl, substituted heteroaryl,        heteroarylalkyl, and substituted heteroarylalkyl, wherein when Y        is nitrogen, R^(d) is not present, or wherein R^(c) and R^(d)        together with Y form a cycloalkyl, substituted cycloalkyl, aryl,        substituted aryl, heteroaryl or substituted heteroaryl group;        and    -   R^(c) is alkyl or substituted alkyl,    -   or salt, solvate or hydrate thereof.

In some instances, R¹ is hydrogen. In some instances, R² is hydrogen. Insome instances, R³ is hydrogen. In some instances, R⁴ is hydrogen. Incertain instances, each of R¹, R², R³ and R⁴ are hydrogen.

In other instances, R¹ is:

-   -   wherein        represents the R¹—O bond;    -   R⁸ is hydrogen, alkyl or substituted alkyl; In some instances,        R⁸ is selected from hydrogen, methyl, ethyl and cyclopropyl.    -   R⁹ is —NR^(f) or —OR^(f), wherein Rr is alkyl, substituted        alkyl, acyl, alkylacyl or substituted alkylacyl, or wherein        R^(f) together with R¹⁰ form a cycloalkyl, substituted        cycloalkyl, heterocycloalkyl, substituted heterocycloalkyl,        aryl, substituted aryl, arylalkyl, substituted arylalkyl,        heteroaryl, substituted heteroaryl, heteroarylalkyl, and        substituted heteroarylalkyl; and    -   R¹⁰ is cycloalkyl, substituted cycloalkyl, heterocycloalkyl,        substituted heterocycloalkyl, aryl, substituted aryl, arylalkyl,        substituted arylalkyl, heteroaryl, substituted heteroaryl,        heteroarylalkyl, and substituted heteroarylalkyl or wherein R¹⁰        together with R^(f) form a cycloalkyl, substituted cycloalkyl,        heterocycloalkyl, substituted heterocycloalkyl, aryl,        substituted aryl, arylalkyl, substituted arylalkyl, heteroaryl,        substituted heteroaryl, heteroarylalkyl, and substituted        heteroarylalkyl.

In certain instances, R¹ is selected from.

wherein

represents the R¹—O bond;

-   -   R⁸ is hydrogen, alkyl, substituted alkyl; and    -   R^(g) is hydrogen or a halogen selected from fluorine, chlorine,        bromine or iodine.

In other instances, R¹ is:

-   -   wherein:    -   R¹¹ is alkyl or substituted alkyl;    -   R¹² is alkyl or alkyl substituted with a cycloalkyl, substituted        cycloalkyl, heterocycloalkyl, substituted heterocycloalkyl,        aryl, substituted aryl, arylalkyl, substituted arylalkyl,        heteroaryl, substituted heteroaryl, heteroarylalkyl, and        substituted heteroarylalkyl.

In some instances, R¹¹ is selected from methyl, ethyl, n-propyl,iso-propyl, n-butyl, iso-butyl, t-butyl. In certain instances, R¹¹ isiso-propyl.

In certain instances, R¹ is:

-   -   wherein R^(h) and R^(i) are each independently selected from        hydrogen, hydroxyl or halogen. In some instances, R^(h) is        hydroxyl. In some instances, R^(h) is F. In some instances,        R^(h) is Cl. In some instances, R^(h) is I. In some instances,        R^(h) is Br. In some instances, R^(i) is hydroxyl. In some        instances, R^(i) is F. In other instances, R^(i) is Cl. In other        instances, R^(i) is I. In other instances, R^(i) is Br.

Formula DCD-(IIA-2)

Aspects of the present disclosure according to certain embodimentsinclude a compound of formula DCD-(IIA-2):

-   -   wherein X is —NHCO— or oxygen;    -   R¹, R² and R³ are each independently selected from hydrogen,        alkyl, substituted alkyl, heteroalkyl, substituted heteroalkyl,        cycloalkyl, substituted cycloalkyl, heterocycloalkyl,        substituted heterocycloalkyl, aryl, substituted aryl, arylalkyl,        substituted arylalkyl, heteroaryl, substituted heteroaryl,        heteroarylalkyl, and substituted heteroarylalkyl;    -   X₁, X₂, X₃, X₄ and X₅ are each independently selected from        carbon, nitrogen, oxygen or sulfur;    -   R^(a) is optionally absent or when present is selected from        hydrogen or oxygen;    -   wherein        indicates a double or single bond;    -   n is an integer from 2 to 25.    -   Y is selected from carbon, nitrogen or silicon;    -   R^(b), R^(c) and R^(d) are independently selected hydrogen,        alkyl, substituted alkyl, heteroalkyl, substituted heteroalkyl,        cycloalkyl, substituted cycloalkyl, heterocycloalkyl,        substituted heterocycloalkyl, aryl, substituted aryl, arylalkyl,        substituted arylalkyl, heteroaryl, substituted heteroaryl,        heteroarylalkyl, and substituted heteroarylalkyl, wherein when Y        is nitrogen, R^(d) is not present, or wherein R^(c) and R^(d)        together with Y form a cycloalkyl, substituted cycloalkyl, aryl,        substituted aryl, heteroaryl or substituted heteroaryl group;        and    -   R^(e) is alkyl or substituted alkyl,    -   or salt, solvate or hydrate thereof.

In some instances, R¹ is hydrogen. In some instances, R² is hydrogen. Insome instances, R³ is hydrogen. In certain instances, each of R¹, R² andR³ are hydrogen. In other instances, R¹ is:

-   -   wherein        represents the R¹—O bond;    -   R⁸ is hydrogen, alkyl or substituted alkyl. In some instances,        R⁸ is selected from hydrogen, methyl, ethyl and cyclopropyl.    -   R⁹ is —NR^(f) or —OR^(f), wherein R^(f) is alkyl, substituted        alkyl, acyl, alkylacyl or substituted alkylacyl, or wherein        R^(f) together with R¹⁰ form a cycloalkyl, substituted        cycloalkyl, heterocycloalkyl, substituted heterocycloalkyl,        aryl, substituted aryl, arylalkyl, substituted arylalkyl,        heteroaryl, substituted heteroaryl, heteroarylalkyl, and        substituted heteroarylalkyl; and    -   R¹⁰ is cycloalkyl, substituted cycloalkyl, heterocycloalkyl,        substituted heterocycloalkyl, aryl, substituted aryl, arylalkyl,        substituted arylalkyl, heteroaryl, substituted heteroaryl,        heteroarylalkyl, and substituted heteroarylalkyl or wherein R¹⁰        together with R^(f) form a cycloalkyl, substituted cycloalkyl,        heterocycloalkyl, substituted heterocycloalkyl, aryl,        substituted aryl, arylalkyl, substituted arylalkyl, heteroaryl,        substituted heteroaryl, heteroarylalkyl, and substituted        heteroarylalkyl.

In certain instances, R¹ is selected from:

-   -   wherein        represents the R¹—O bond,    -   R⁸ is hydrogen, alkyl, substituted alkyl; and    -   R^(g) is hydrogen or a halogen selected from fluorine, chlorine,        bromine or iodine.

In other instances, R¹ is:

-   -   wherein:    -   R¹¹ is alkyl or substituted alkyl;    -   R¹² is alkyl or alkyl substituted with a cycloalkyl, substituted        cycloalkyl, heterocycloalkyl, substituted heterocycloalkyl,        aryl, substituted aryl, arylalkyl, substituted arylalkyl,        heteroaryl, substituted heteroaryl, heteroarylalkyl, and        substituted heteroarylalkyl.

In some instances, R¹¹ is selected from methyl, ethyl, n-propyl,iso-propyl, n-butyl, iso-butyl, t-butyl. In certain instances, R¹¹ isiso-propyl.

In certain instances, R¹ is:

-   -   wherein R^(h) and R^(i) are each independently selected from        hydrogen, hydroxyl or halogen. In some instances, R^(h) is        hydroxyl. In some instances, R^(h) is F. In some instances,        R^(h) is Cl. In some instances, R^(h) is I. In some instances,        R^(a) is Br. In some instances, R^(i) is hydroxyl. In some        instances, R^(i) is F. In other instances, R^(i) is Cl. In other        instances, R^(i) is I.

In other instances, R^(i) is Br.

Formula DCD-(IIA-3)

Aspects of the present disclosure according to certain embodimentsinclude a compound of formula DCD-(IIA-3):

-   -   wherein X is —NHCO— or oxygen.    -   R¹, R², R³ and R⁴ are each independently selected from hydrogen,        alkyl, substituted alkyl, heteroalkyl, substituted heteroalkyl,        cycloalkyl, substituted cycloalkyl, heterocycloalkyl,        substituted heterocycloalkyl, aryl, substituted aryl, arylalkyl,        substituted arylalkyl, heteroaryl, substituted heteroaryl,        heteroarylalkyl, and substituted heteroarylalkyl;    -   X₁, X₂, X₃, X₄ and X₅ are each independently selected from        carbon, nitrogen, oxygen or sulfur;    -   R is optionally absent or wen present is selected from hydrogen        or oxygen;    -   wherein        a indicates a double or single bond;    -   n is an integer from 2 to 25;    -   Y is selected from carbon, nitrogen or silicon;    -   R^(b), R^(c) and R^(d) are independently selected hydrogen,        alkyl, substituted alkyl, heteroalkyl, substituted heteroalkyl,        cycloalkyl, substituted cycloalkyl, heterocycloalkyl,        substituted heterocycloalkyl, aryl, substituted aryl, arylalkyl,        substituted arylalkyl, heteroaryl, substituted heteroaryl,        heteroarylalkyl, and substituted heteroarylalkyl, wherein when Y        is nitrogen, R^(d) is not present, or wherein R^(c) and R^(d)        together with Y form a cycloalkyl, substituted cycloalkyl, aryl,        substituted aryl, heteroaryl or substituted heteroaryl group;        and    -   R^(e) is alkyl or substituted alkyl,    -   or salt, solvate or hydrate thereof.

In some instances, R¹ is hydrogen. In some instances, R² is hydrogen. Insome instances, R³ is hydrogen. In some instances, R⁴ is hydrogen. Incertain instances, each of R¹, R², R³ and R⁴ are hydrogen.

In other instances, R¹ is:

-   -   wherein        represents the R¹—O bond;    -   R⁸ is hydrogen, alkyl or substituted alkyl; In some instances,        R⁸ is selected from hydrogen, methyl, ethyl and cyclopropyl.    -   R⁹ is —NR^(f) or —OR^(f), wherein R^(f) is alkyl, substituted        alkyl, acyl, alkylacyl or substituted alkylacyl, or wherein Rr        together with R¹⁰ form a cycloalkyl, substituted cycloalkyl,        heterocycloalkyl, substituted heterocycloalkyl, aryl,        substituted aryl, arylalkyl, substituted arylalkyl, heteroaryl,        substituted heteroaryl, heteroarylalkyl, and substituted        heteroarylalkyl; and    -   R¹⁰ is cycloalkyl, substituted cycloalkyl, heterocycloalkyl,        substituted heterocycloalkyl, aryl, substituted aryl, arylalkyl,        substituted arylalkyl, heteroaryl, substituted heteroaryl,        heteroarylalkyl, and substituted heteroarylalkyl or wherein R¹⁰        together with R^(f) form a cycloalkyl, substituted cycloalkyl,        heterocycloalkyl, substituted heterocycloalkyl, aryl,        substituted aryl, arylalkyl, substituted arylalkyl, heteroaryl,        substituted heteroaryl, heteroarylalkyl, and substituted        heteroarylalkyl.

In certain instances, R¹ is selected from:

-   -   wherein        represents the R¹—O bond;    -   R⁸ is hydrogen, alkyl, substituted alkyl; and    -   R^(g) is hydrogen or a halogen selected from fluorine, chlorine,        bromine or iodine.    -   In other instances, R¹ is:

-   -   wherein:    -   R¹¹ is alkyl or substituted alkyl;    -   R¹² is alkyl or alkyl substituted with a cycloalkyl, substituted        cycloalkyl, heterocycloalkyl, substituted heterocycloalkyl,        aryl, substituted aryl, arylalkyl, substituted arylalkyl,        heteroaryl, substituted heteroaryl, heteroarylalkyl, and        substituted heteroarylalkyl.

In some instances, R¹¹ is selected from methyl, ethyl, n-propyl,iso-propyl, n-butyl, iso-butyl, t-butyl. In certain instances, R¹¹ isiso-propyl.

In certain instances, R¹ is:

-   -   wherein R^(h) and R^(i) are each independently selected from        hydrogen, hydroxyl or halogen. In some instances, R^(h) is        hydroxyl. In some instances, R^(h) is F. In some instances, R is        Cl. In some instances, R^(h) is I. In some instances, R^(h) is        Br. In some instances, R^(i) is hydroxyl. In some instances,        R^(i) is F. In other instances, R^(i) is Cl. In other instances,        R^(i) is I. In other instances, R^(i) is Br.

Formula DCD-(IIA-4)

Aspects of the present disclosure according to certain embodimentsinclude a compound of formula DCD-(IIA-4):

-   -   wherein X is —NHCO— or oxygen;    -   R¹ and R² are each independently selected from hydrogen, alkyl,        substituted alkyl, heteroalkyl, substituted heteroalkyl,        cycloalkyl, substituted cycloalkyl, heterocycloalkyl,        substituted heterocycloalkyl, aryl, substituted aryl, arylalkyl,        substituted arylalkyl, heteroaryl, substituted heteroaryl,        heteroarylalkyl, and substituted heteroarylalkyl;    -   X₁, X₂. X₃, X₄ and X₅ are each independently selected from        carbon, nitrogen, oxygen or sulfur;    -   R¹ is optionally absent or when present is selected from        hydrogen or oxygen;    -   wherein        indicates a double or single bond;    -   n is an integer from 2 to 25;    -   Y is selected from carbon, nitrogen or silicon;    -   R^(b), R^(c) and R^(d) are independently selected hydrogen,        alkyl, substituted alkyl, heteroalkyl, substituted heteroalkyl,        cycloalkyl, substituted cycloalkyl, heterocycloalkyl,        substituted heterocycloalkyl, amyl, substituted aryl, arylalkyl,        substituted arylalkyl, heteroaryl, substituted heteroaryl,        heteroarylalkyl, and substituted heteroarylalkyl, wherein when Y        is nitrogen, R^(d) is not present, or wherein R^(c) and R^(d)        together with Y form a cycloalkyl, substituted cycloalkyl, aryl,        substituted aryl, heteroaryl or substituted heteroaryl group;        and    -   R^(e) is alkyl or substituted alkyl,    -   or salt, solvate or hydrate thereof.

In some instances, R¹ is hydrogen. In some instances, R² is hydrogen. Incertain instances, each of R¹ and R² are hydrogen.

In other instances, R¹ is:

-   -   wherein        represents the R¹—O bond;    -   R⁸ is hydrogen, alkyl or substituted alkyl; In some instances,        R⁸ is selected from hydrogen, methyl, ethyl and cyclopropyl.    -   R⁹ is —NR^(f) or —OR wherein R^(f) is alkyl, substituted alkyl,        acyl, alkylacyl or substituted alkylacyl, or wherein R^(f)        together with R¹⁰ form a cycloalkyl, substituted cycloalkyl,        heterocycloalkyl, substituted heterocycloalkyl, aryl,        substituted aryl, arylalkyl, substituted arylalkyl, heteroaryl,        substituted heteroaryl, heteroarylalkyl, and substituted        heteroarylalkyl; and    -   R¹⁰ is cycloalkyl, substituted cycloalkyl, heterocycloalkyl,        substituted heterocycloalkyl, aryl, substituted aryl, arylalkyl,        substituted arylalkyl, heteroaryl, substituted heteroaryl,        heteroarylalkyl, and substituted heteroarylalkyl or wherein R¹⁰        together with R^(f) form a cycloalkyl, substituted cycloalkyl,        heterocycloalkyl, substituted heterocycloalkyl, aryl,        substituted aryl, arylalkyl, substituted arylalkyl, heteroaryl,        substituted heteroaryl, heteroarylalkyl, and substituted        heteroarylalkyl.

In certain instances, R¹ is selected from:

-   -   wherein        represents the R¹—O bond;    -   R⁸ is hydrogen, alkyl, substituted alkyl; and    -   R^(g) is hydrogen or a halogen selected from fluorine, chlorine,        bromine or iodine.

In other instances, R¹ is:

-   -   wherein:

-   -   R¹¹ is alkyl or substituted alkyl.    -   R¹² is alkyl or alkyl substituted with a cycloalkyl, substituted        cycloalkyl, heterocycloalkyl, substituted heterocycloalkyl,        aryl, substituted aryl, arylalkyl, substituted arylalkyl,        heteroaryl, substituted heteroaryl, heteroarylalkyl, and        substituted heteroarylalkyl.

In some instances, R¹¹ is selected from methyl, ethyl, n-propyl,iso-propyl, n-butyl, iso-butyl, t-butyl. In certain instances, R¹¹ isiso-propyl.

In certain instances, R¹ is:

-   -   wherein R^(h) and R^(i) are each independently selected from        hydrogen, hydroxyl or halogen. In some instances, R^(h) is        hydroxyl. In some instances, R^(h) is F. In some instances,        R^(h) is Cl. In some instances, R^(h) is I. In some instances,        R^(h) is Br. In some instances, R^(i) is hydroxyl. In some        instances, R^(i) is F. In other instances, R^(i) is Cl. In other        instances, R^(i) is I.

In other instances, R^(i) is

Formula DCD-(IIB-1)

Aspects of the present disclosure according to certain embodimentsinclude a compound of formula DCD-(IIB-1):

-   -   wherein:    -   Z is selected from:

-   -   wherein        indicates the Z—O bond;    -   wherein X is —NHCO— or oxygen;    -   R¹, R², R³ and R⁴ are each independently selected from hydrogen,        alkyl, substituted alkyl, heteroalkyl, substituted heteroalkyl,        cycloalkyl, substituted cycloalkyl, heterocycloalkyl,        substituted heterocycloalkyl, aryl, substituted aryl, arylalkyl,        substituted arylalkyl, heteroaryl, substituted heteroaryl,        heteroarylalkyl, and substituted heteroarylalkyl;    -   n is an integer from 2 to 25;    -   Y is selected from carbon, nitrogen or silicon;    -   R^(b), R^(c) and R^(d) are independently selected hydrogen,        alkyl, substituted alkyl, heteroalkyl, substituted heteroalkyl,        cycloalkyl, substituted cycloalkyl, heterocycloalkyl,        substituted heterocycloalkyl, aryl, substituted aryl, arylalkyl,        substituted arylalkyl, heteroaryl, substituted heteroaryl,        heteroarylalkyl, and substituted heteroarylalkyl, wherein when Y        is nitrogen, R^(d) is not present, or wherein R^(c) and R^(d)        together with Y form a cycloalkyl, substituted cycloalkyl, aryl,        substituted aryl, heteroaryl or substituted heteroaryl group;        and    -   R^(e) is alkyl or substituted alkyl,    -   or salt, solvate or hydrate thereof.

Formula DCD-(IIB-2)

Aspects of the present disclosure according to certain embodimentsinclude a compound of formula DCD-(IIB-2):

-   -   wherein:    -   Z is selected from:

wherein

indicates the Z—O bond

-   -   wherein X is —NHCO— or oxygen;    -   R¹, R², R³ and R⁴ are each independently selected from hydrogen,        alkyl, substituted alkyl, heteroalkyl, substituted heteroalkyl,        cycloalkyl, substituted cycloalkyl, heterocycloalkyl,        substituted heterocycloalkyl, aryl, substituted aryl, arylalkyl,        substituted arylalkyl, heteroaryl, substituted heteroaryl,        heteroarylalkyl, and substituted heteroarylalkyl;    -   n is an integer from 2 to 25;    -   Y is selected from carbon, nitrogen or silicon;    -   R^(b), R^(c) and R^(d) are independently selected hydrogen,        alkyl, substituted alkyl, heteroalkyl, substituted heteroalkyl,        cycloalkyl, substituted cycloalkyl, heterocycloalkyl,        substituted heterocycloalkyl, aryl, substituted aryl, arylalkyl,        substituted arylalkyl, heteroaryl, substituted heteroaryl,        heteroarylalkyl, and substituted heteroarylalkyl, wherein when Y        is nitrogen, R^(d) is not present, or wherein R^(c) and R^(d)        together with Y form a cycloalkyl, substituted cycloalkyl, aryl,        substituted aryl, heteroaryl or substituted heteroaryl group;        and    -   R^(e) is alkyl or substituted alkyl,    -   or salt, solvate or hydrate thereof.

Formula DCD-(IIB-3)

Aspects of the present disclosure according to certain embodimentsinclude a compound of formula DCD-(IIB-3):

-   -   wherein:    -   Z is selected from:

-   -   wherein        indicates the Z—O bond;    -   wherein X is —NHCO— or oxygen;    -   R¹, R², R³ and R⁴ are each independently selected from hydrogen,        alkyl, substituted alkyl, heteroalkyl, substituted heteroalkyl,        cycloalkyl, substituted cycloalkyl, heterocycloalkyl,        substituted heterocycloalkyl, aryl, substituted aryl, arylalkyl,        substituted arylalkyl, heteroaryl, substituted heteroaryl,        heteroarylalkyl, and substituted heteroarylalkyl;    -   n is an integer from 2 to 25;    -   Y is selected from carbon, nitrogen or silicon,    -   R^(b), R^(c) and R^(d) are independently selected hydrogen,        alkyl, substituted alkyl, heteroalkyl, substituted heteroalkyl,        cycloalkyl, substituted cycloalkyl, heterocycloalkyl,        substituted heterocycloalkyl, aryl, substituted aryl, arylalkyl,        substituted arylalkyl, heteroaryl, substituted heteroaryl,        heteroarylalkyl, and substituted heteroarylalkyl, wherein when Y        is nitrogen, R^(d) is not present, or wherein R^(c) and R^(d)        together with Y form a cycloalkyl, substituted cycloalkyl, aryl,        substituted aryl, heteroaryl or substituted heteroaryl group;        and    -   R^(e) is alkyl or substituted alkyl.    -   or salt, solvate or hydrate thereof.

In embodiments of compounds of formulae DCD-(II), DCD-(IIA) andDCD-(IIB), in some instances, R^(e) may be alkyl or substituted alkyl.In some instances, R^(e) is a C8 to C20 alkyl. In some instances, R^(e)is a substituted C8 to C20 alkyl. In certain instances, R^(e) is a C13alkyl.

In some instances n is an integer from 2 to 25, such as where n isselected from 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17,18, 19, 20, 21, 22, 23, 24 and 25.

In some instances, Y is carbon. In some instances, Y is nitrogen. Insome instances, Y is silicon.

In some instances, R^(b) is hydrogen. In some instances, R^(b) is alkyl.In some instances, R^(b) is a C1-C16 alkyl. In some instances, R^(b) isselected from methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl,tert-butyl, pentyl, hexyl, heptyl and octyl. In some instances, R^(b) isselected from:

-   -   wherein        indicates a bond to Y and R^(z) is hydrogen, alkyl or alkyl        substituted with a cycloalkyl, substituted cycloalkyl,        heterocycloalkyl, substituted heterocycloalkyl, aryl,        substituted aryl, arylalkyl, substituted arylalkyl, heteroaryl,        substituted heteroaryl, heteroarylalkyl, and substituted        heteroarylalkyl. In certain instances, R^(z) is alkyl, such as a        C1 to C16 alkyl or C1 to C16 substituted alkyl.

In some instances, R^(c) is hydrogen. In some instances, R^(c) is alkyl.In some instances, R^(c) is a C1-C16 alkyl. In some instances, R^(c) isselected from methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl,tert-butyl, pentyl, hexyl, heptyl and octyl.

In some embodiments, R^(d) is a C5 to C25 alkyl or a C5 to C25 alkylsubstituted with a cycloalkyl group, substituted cycloalkyl group,heterocycloalkyl group, substituted heterocycloalkyl group, aryl group,substituted aryl group, arylalkyl group, substituted arylalkyl group,heteroaryl group, substituted heteroaryl group, heteroarylalkyl group,or substituted heteroarylalkyl group. In some instances, R^(d) is a C5to C25 alkyl substituted with a moiety selected from the groupconsisting of:

-   -   wherein        indicates the bond to the C5 to C25 alkyl; and R^(m) and R^(n)        are independently selected from hydrogen, halogen, hydroxyl,        substituted hydroxyl, amino, substituted amino, thiol,        substituted thiol, sulfoxide, substituted sulfoxide, sulfone,        substituted sulfone, sulfoximine, substituted sulfoximine, acyl,        aminoacyl, alkyl, substituted alkyl, heteroalkyl, substituted        heteroalkyl, cycloalkyl, substituted cycloalkyl, spiroalkyl,        heterocycloalkyl, substituted heterocycloalkyl, aryl,        substituted aryl, arylalkyl, substituted arylalkyl, heteroaryl,        substituted heteroaryl, heteroarylalkyl, and substituted        heteroarylalkyl.

In some instances, R^(m) is hydrogen. In some instances, R^(m) ishalogen. In some instances, R^(m) is selected from fluorine, bromine oriodine. In some instances, R^(n) is hydrogen. In some instances, R^(n)is halogen. In some instances, R^(n) is fluorine, bromine or iodine.

In some embodiments, compounds of interest include those shown in Tables1-9, which are not to be construed as limitative.

TABLE 1

TABLE 2 Amide Analogs

TABLE 3 Trifluoromethyl Aminomethyl Analogs

TABLE 4 Sulfoximine Analogs

TABLE 5 Vinylfluoride Analogs

TABLE 6 Aminooxetane Analogs

TABLE 7 Pyrazole Analoge

TABLE 8 Tetrazolone Analogs

TABLE 9 Imidazole Analogs

Methods for Activating iNKT Cells and Selectively Eliminating SenescentCells

As summarized above, aspects of the present disclosure also includemethods for activating an iNKT cell. In embodiments, methods includecontacting an iNKT cell with an amount of one or more of the compoundsor a pharmaceutically acceptable salt thereof described hereinsufficient to activate the iNKT cell. In some instances, a source of theiNKT cell is contacted in vitro. In other instances, a source of theiNKT cell is contacted in vivo (e.g., by administering to a subject asdescribed in greater detail below). In still other instances, a sourceof the iNKT cell is contacted ex vivo.

In some embodiments, methods include contacting one or more of thecompounds described herein with iNKT cells in a manner sufficient toactivate the iNKT cells, where the activated iNKT cells induce aTH1-type cytokine response (e.g., increase production of one or morecytokines selected from IFN-γ, IL-1β, IL-2, IL-3, IL-8, IL-12, IL-15,TNF-α, GM-CSF, RANTES, MIP-1α and MCP-1). In other instances, theactivated iNKT cells induce a T_(H)2-type cytokine response (e.g.,increase production of one or more cytokines selected from IL-4, IL-6,IL-8, IL-10, IL-13, RANTES, MIP-1α and MCP-1). In some instances,activating the iNKT cells with one or more of the subject compounds issufficient to increase cytokine production by 1% or more as compared toa suitable control (e.g., iNKT cells not contacted with the compound ora control compound), such as by 2% or more, such as by 3% or more, suchas by 4% or more, such as by 5% or more, such as by 10% or more, such asby 15% or more, such as by 20% or more, such as by 25% or more, such asby 50% or more, such as by 75% or more, such as by 90% or more, such asby 95% or more and including by 99% or more.

In certain instances, activating the iNKT cells with one or more of thesubject compounds is sufficient to increase interleukin-2 (IL-2)production by 1% or more as compared to a suitable control (e.g., iNKTcells not contacted with the compound or a control compound), such as by2% or more, such as by 3% or more, such as by 4% or more, such as by 5%or more, such as by 10% or more, such as by 15% or more, such as by 20%or more, such as by 25% or more, such as by 50% or more, such as by 75%or more, such as by 90% or more, such as by 95% or more and including by99% or more. In certain instances, activating the iNKT cells with one ormore of the subject compounds is sufficient to increase interleukin-4(IL-4) production by 1% or more as compared to a suitable control (e.g.,iNKT cells not contacted with the compound or a control compound), suchas by 2% or more, such as by 3% or more, such as by 4% or more, such asby 5% or more, such as by 10% or more, such as by 15% or more, such asby 20% or more, such as by 25% or more, such as by 50% or more, such asby 75% or more, such as by 90% or more, such as by 95% or more andincluding by 99% or more. In certain instances, activating the iNKTcells with one or more of the subject compounds is sufficient toincrease interleukin-6 (IL-6) production by 1% or more as compared to asuitable control (e.g., iNKT cells not contacted with the compound or acontrol compound), such as by 2% or more, such as by 3% or more, such asby 4% or more, such as by 5% or more, such as by 10% or more, such as by15% or more, such as by 20% or more, such as by 25% or more, such as by50% or more, such as by 75% or more, such as by 90% or more, such as by95% or more and including by 99% or more. In certain instances,activating the iNKT cells with one or more of the subject compounds issufficient to increase interferon gamma (IFNγ) production by 1% or moreas compared to a suitable control (e.g., iNKT cells not contacted withthe compound or a control compound), such as by 2% or more, such as by3% or more, such as by 4% or more, such as by 5% or more, such as by 10%or more, such as by 15% or more, such as by 20% or more, such as by 25%or more, such as by 50% or more, such as by 75% or more, such as by 90%or more, such as by 95% or more and including by 99% or more. In certaininstances, activating the iNKT cells with one or more of the subjectcompounds is sufficient to increase tumor necrosis factor (TNFα)production by 1% or more as compared to a suitable control (e.g., iNKTcells not contacted with the compound or a control compound), such as by2% or more, such as by 3% or more, such as by 4% or more, such as by 5%or more, such as by 10% or more, such as by 15% or more, such as by 20%or more, such as by 25% or more, such as by 50% or more, such as by 75%or more, such as by 90% or more, such as by 95% or more and including by99% or more.

In certain instances, activating the iNKT cells with one or more of thesubject compounds is sufficient to increase cytokine production ascompared to contacting the iNKT cells with α-galactosylceramide(α-GalCer), such as where cytokine production (e.g., increasing one ormore of IFN-γ, TL-1β, IL-2, IL-3, IL-8, IL-12, IL-15, TNF-α, GM-CSF,RANTES, MIP-1α and MCP-1 or IL-4, IL-6, IL-8, IL-10, IL-13, RANTES,MIP-1α and MCP-1) is higher by activating the iNKT cells with one ormore of the subject compounds than when the iNKT cells are contactedwith α-galactosylceramide. In some embodiments, the compounds of thepresent disclosure increase cytokine production by 1% or more ascompared to α-galactosylceramide, such as by 2% or more, such as by 3%or more, such as by 4% or more, such as by 5% or more, such as by 10% ormore, such as by 15% or more, such as by 20% or more, such as by 25% ormore, such as by 50% or more, such as by 75% or more, such as by 90% ormore, such as by 95% or more and including by 99% or more as compared toα-galactosylceramide.

In some instances, the compound forms a complex with a CD1 molecule onan antigen-presenting cell. In certain instances, the CD1 molecule is aCD1d molecule. In some instances, the receptor on the T lymphocyte is aT cell receptor. In some instances, the compound stimulates at least oneother lymphocyte to produce the cytokine response in some instances theat least one other lymphocyte is a T helper cell. In some embodiments,methods include activating iNKT cells with the subject compounds in amanner sufficient to modulate an immune response in a subject.

In practicing the subject methods, the iNKT cells may be contacted withthe subject compounds for a duration of 1 minute or more, such as for 2minutes or more, such as for 3 minutes or more, such as for 4 minutes ormore, such as for 5 minutes or more, such as for 10 minutes or more,such as for 15 minutes or more, such as for 30 minutes or more, such asfor 60 minutes or more, such as for 2 hours or more, such as for 6 hoursor more, such as for 12 hours or more, such as for 18 hours or more andincluding for 24 hours or more. In certain embodiments, the productionof one or more cytokines may be assessed (e.g., quantified) aftercontacting the compound with the iNKT cells. In some instances, theproduction of cytokines is assessed in real time (i.e., continuouslymonitored). In other instances, the production of cytokines is assessedat predetermined time intervals, such as every 1 minute, every 15minutes, every 30 minutes, every 60 minutes, every 2 hours, every 4hours, every 6 hours, every 12 hours, every 18 hours, including every 24hours.

In some embodiments, contacting iNKT cells with one or more of thecompounds of the present disclosure is sufficient to activate iNKT cellsand to reduce the presence of or induce the killing of senescent cells.In certain embodiments, the senescent cells are senescent cells havingan inflammatory secretome. For example, activating iNKT cells with thesubject compounds according to these embodiments produces a cytotoxiceffect against senescent cells. In some instances, methods includeactivating iNKT cells with the subject compounds in a manner sufficientto reduce the presence of or induce the killing of senescent cells invitro. In some instances, methods include activating iNKT cells with thesubject compounds in a manner sufficient to reduce the presence of orinduce the killing of senescent cells in vivo (such as by administeringthe compound to a subject as part of a pharmaceutical compositiondescribed below). In some instances, activating iNKT cells with thesubject compounds is sufficient to reduce the presence of senescentcells by 1% or more, such as by 2% or more, such as by 3% or more, suchas by 4% or more, such as by 5% or more, such as by 10% or more, such asby 15% or more, such as by 20% or more, such as by 25% or more, such asby 50% or more, such as by 75% or more, such as by 90% or more, such asby 95% or more and including by 99% or more. In certain instances, thesubject compounds eliminate the presence of senescent cells (e.g., whereactivation of iNKT cells reduces senescent present by 100%). Thereduction in the presence of senescent cells may be assessed (e.g.,quantified) after contacting the compound with the iNKT cells.

In some embodiments, iNKT cells activated by contact with the compoundsdescribed herein selectively reduce the presence of or selectivelyinduce killing of senescent cells while maintaining (i.e., not killing)healthy cells. In some instances, contacting iNKT cells activated withthe compounds of the present disclosure is sufficient to reduce thepresence of senescent cells while maintaining 75% or more of the healthycells, such as 80% or more, such as 85% or more, such as 90% or more,such as 95% or more, such as 97% or more, such as 99% or more, such as99.9% or more and including 99.99% or more. In certain instances, iNKTcells activated by contact with compounds of the present disclosureselectively reduce the presence of senescent cells without any effect onhealthy cells (i.e., 100% of healthy cells are maintained).

In some instances, the reduction of senescent cells may be assessed inreal time (i.e., continuously monitored). In other instances, thereduction of senescent cells is assessed at predetermined timeintervals, such as every 1 minute, every 15 minutes, every 30 minutes,every 60 minutes, every 2 hours, every 4 hours, every 6 hours, every 12hours, every 18 hours, including every 24 hours.

Aspects of the present disclosure also include administering one or moreof the compounds described herein to a subject in need thereof. Inembodiments, the term “subject” is meant the person or organism to whichthe compound is administered. As such, subjects of the presentdisclosure may include but are not limited to mammals, e.g., humans andother primates, such as chimpanzees and other apes and monkey species,dogs, rabbits, cats and other domesticated pets; and the like, where incertain embodiments the subject are humans. The term “subject” is alsomeant to include a person or organism of any age, weight or otherphysical characteristic, where the subjects may be an adult, a child, aninfant or a newborn.

In certain embodiments, the subject is diagnosed as having an autoimmunedisease, fibrotic disorders (lung, kidney, liver), an allergic disease,a metabolic syndrome, type 2 diabetes. NAFLD. NASH, cancer, pathogeninfection, rheumatoid arthritis, ulcerative colitis, multiple sclerosis,familial hypercholesteremia, giant cell arteritis, idiopathic pulmonaryfibrosis, systemic lupus erythematosus, cachexia, glaucoma, chronicobstructive pulmonary disease, systemic sclerosis, pulmonary arterialhypertension, lipodystrophy, sarcopenia, alopecia, post myocardialinfarction, vitiligo, POTS, MCAD, Sjogren's, Scleroderma, HashimotoDisease, Ankylosing Spondylitis, Fibromyalgia, Sarcoidosis, Hepatitis,Raynauld's Syndrome, Mold Illness, Celiac, Crohn's, Pemphigus, SPS, PBC,Psoriatic Arthritis, CIDP, motor neuron disease, GPA, ALS, myasenthiagravis, and presbyopia. In some embodiments, the subject is diagnosed(e.g., by clinical laboratory test or by a qualified healthcareprofessional) as having or exhibiting at least one symptom of multiplesclerosis, articular rheumatism, psoriasis, Crohn's disease, leukodermavulgaris, Behcet's disease, collagenosis, type I diabetes mellitus,uveitis, Sjogren's syndrome, autoimmune cardiomyotitis, autoimmune liverdisease, autoimmune gastritis, pemphigus, Guillain-Barre syndrome,HTLV-1-related myelopathy or fulminant hepatitis.

In some embodiments, methods include administering one or more of thecompounds to treat a subject for an infectious disease, such as onecaused by a pathogenic microbe, including for example viruses, bacteria,fungi, protozoa and multicellular parasites. In one example, theinfectious disease is by a virus selected from Retroviridae,Picornaviridae, Caliciviridae, Togaviridae, Flaviridae, Coronaviridae,Rhabdoviridae, Filoviridae, Paramyxoviridae, Orthomyxovindae,Bungaviridae, Arena viridae, Reovindae, Bimaviridae, Hepadnaviridae,Parvoviridae, Papovaviridae, Adenoviridae, Herpesviridae, Poxyiridae andIridoviridae. In another example, the infectious disease is caused by abacteria selected from Helicobacter pylori, Borrelia burgdorferi,Legionella pneumophilia, Klebsiella pneumoniae, Mycobacteria sps,Staphylococcus aureus, Neisseria gonorrhoeae, Neisseria meningitidis,Listeria monocytogenes, Streptococcus pyogenes, Streptococcusagalactiae, Streptococcus, Streptococcus faecalis, Streptococcus bovis,Streptococcus pneumoniae, pathogenic Campylobacter sp., Enterococcussp., Chlamydia sp., Haemophilus influenzae, Bacillus antracis,Corynebacterium diphtheriae, Corynebacterium sp., Erysipelothrixrhusiopathiae, Clostridium perfringens, Clostridium tetani, Enterobacteraerogenes. Klebsiella pneumoniae, Pasteurella multocida, Bacteroidessp., Fusobacterium nucleatum, Streptobacillus moniliformis, Treponemapallidium, Treponema pertenue, Leptospira, Actinomyces israelii,Sphingomonas capsulata and Francisella tularensis.

Compounds as described herein may be administered to a subject by anyconvenient protocol, including, but not limited, to intraperitoneally,topically, orally, sublingually, parenterally, intravenously, vaginally,rectally as well as by transdermal protocols. In certain embodiments,the subject compounds are administered by intravenous injection. Incertain embodiments, the subject compounds are administered byintraperitoneal injection.

Depending on the condition being treated, the amount of compoundadministered to the subject may vary, such as ranging from about 100mg/day to about 10.000 mg/day, such as from about 10 mg/day to about9000 mg/day, such as from 50 mg/day to about 8000 mg/day, such as fromabout 100 mg/day to about 7000 mg/day, such as from about 500 mg/day toabout 6000 mg/day, including from about 600 mg/day to about 5000 mg/day.Each dosage of the compound or pharmaceutically acceptable saltadministered to the subject may vary ranging from about 1 mg/kg to about1000 mg/kg, such as from about 2 mg/kg to about 900 mg/kg, such as fromabout 3 mg/kg to about 800 mg/kg, such as from about 4 mg/kg to about700 mg/kg, such as from 5 mg/kg to about 600 mg/kg, such as from 6 mg/kgto about 500 mg/kg, such as from 7 mg/kg to about 400 mg/kg, such asfrom about 8 mg/kg to about 300 mg/kg, such as from about 9 mg/kg toabout 200 mg/kg and including from about 10 mg/kg to about 100 mg/kg.

In certain embodiments, protocols may include multiple dosage intervals.By “multiple dosage intervals” is meant that two or more dosages of thecompound is administered to the subject in a sequential manner. Inpracticing methods of the present disclosure, treatment regimens mayinclude two or more dosage intervals, such as three or more dosageintervals, such as four or more dosage intervals, such as five or moredosage intervals, including ten or more dosage intervals. The durationbetween dosage intervals in a multiple dosage interval treatmentprotocol may vary, depending on the physiology of the subject or by thetreatment protocol as determined by a health care professional. Forexample, the duration between dosage intervals in a multiple dosagetreatment protocol may be predetermined and follow at regular intervals.As such, the time between dosage intervals may vary and may be 1 day orlonger, such as 2 days or longer, such as 4 days or longer, such as 6days or longer, such as 8 days or longer, such as 12 days or longer,such as 16 days or longer and including 24 days or longer. In certainembodiments, multiple dosage interval protocols provide for a timebetween dosage intervals of 1 week or longer, such as 2 weeks or longer,such as 3 weeks or longer, such as 4 weeks or longer, such as 5 weeks orlonger, including 6 weeks or longer.

The cycles of drug administration may be repeated for 1, 2, 3, 4, 5, 6,7, 8 or more than 8 dosage cycles, for a total period of 6 months or 1year or 2 years or 3 years or 4 years or more. In certain embodiments,one or more of the subject compounds are administered for the rest ofthe subject's lifetime.

In certain embodiments, compounds of the present disclosure can beadministered prior to, concurrent with, or subsequent to othertherapeutic agents for treating the same or an unrelated condition. Ifprovided at the same time as another therapeutic agent, the presentcompounds may be administered in the same or in a different composition.Thus, the compounds of interest and other therapeutic agents can beadministered to the subject by wav of concurrent therapy. By “concurrenttherapy” is intended administration to a subject such that thetherapeutic effect of the combination of the substances is caused in thesubject undergoing therapy. For example, concurrent therapy may beachieved by administering the compounds of the present disclosure with apharmaceutical composition having at least one other agent, such as ananti-inflammatory agent, immunosuppressant, steroid, analgesic,anesthetic, antihypertensive, chemotherapeutic, among other types oftherapeutics, which in combination make up a therapeutically effectivedose, according to a particular dosing regimen. Administration of theseparate pharmaceutical compositions can be performed simultaneously orat different times (i.e., sequentially, in either order, on the sameday, or on different days), so long as the therapeutic effect of thecombination of these substances is caused in the subject undergoingtherapy.

Where the compounds of the present disclosure is administeredconcurrently with a second therapeutic agent to treat the same condition(e.g., a chemotherapeutic, an anti-viral drug, etc.) the weight ratio ofthe subject compound to second therapeutic agent may range from 1:2 and1:2.5; 1:2.5 and 1:3; 1:3 and 1:3.5 1:3.5 and 1:4; 1:4 and 1.4.5; 1.4.5and 1:5; 1:5 and 1:10, and 1:10 and 1:25 or a range thereof. Forexample, the weight ratio of the subject compound to second therapeuticagent may range between 1.1 and 1:5; 1:5 and 1:10, 1:10 and 1.15; or1:15 and 1:25. Alternatively, the weight ratio of the second therapeuticagent to the subject compound ranges between 2:1 and 2.5:1; 2.5:1 and3:1; 3:1 and 3.5:1; 3.5:1 and 4:1; 4:1 and 4.5:1; 4.5:1 and 5:1; 5:1 and10.1; and 10:1 and 25.1 or a range thereof. For example, the ratio ofthe second therapeutic agent the subject compound may range between 1:1and 5:1, 5:1 and 10:1; 10:1 and 15:1; or 15:1 and 25:1.

Aspects of the present disclosure also include compositions having apharmaceutically acceptable carrier and one or more of the compoundsdescribed above. A wide variety of pharmaceutically acceptableexcipients is known in the art and need not be discussed in detailherein. Pharmaceutically acceptable excipients have been amply describedin a variety of publications, including, for example, A. Gennaro (2000)“Remington: The Science and Practice of Pharmacy”, 20th edition,Lippincott, Williams. & Wilkins; Pharmaceutical Dosage Forms and DrugDelivery Systems (1999) H. C. Ansel et al., eds 7th ed., Lippincott,Williams, & Wilkins; and Handbook of Pharmaceutical Excipients (2000) A.H. Kibbe et al., eds., 3rd ed. Amer. Pharmaceutical Assoc. For example,the one or more excipients may include sucrose, starch, mannitol,sorbitol, lactose, glucose, cellulose, talc, calcium phosphate orcalcium carbonate, a binder (e.g., cellulose, methylcellulose,hydroxymethylcellulose, polypropylpyrrolidone, polyvinylpyrrolidone,gelatin, gum arabic, poly(ethylene glycol), sucrose or starch), adisintegrator (e.g., starch, carboxymethylcellulose, hydroxypropylstarch, low substituted hydroxypropylcellulose, sodium bicarbonate,calcium phosphate or calcium citrate), a lubricant (e.g., magnesiumstearate, light anhydrous silicic acid, talc or sodium lauryl sulfate),a flavoring agent (e.g., citric acid, menthol, glycine or orange pow %der), a preservative (e.g., sodium benzoate, sodium bisulfite,methylparaben or propylparaben), a stabilizer (e.g., citric acid, sodiumcitrate or acetic acid), a suspending agent (e.g., methylcellulose,polyvinylpyrrolidone or aluminum stearate), a dispersing agent (e.g.,hydroxy propylmethylcellulose), a diluent (e.g., water), and base wax(e.g., cocoa butter, white petrolatum or polyethylene glycol).

The compounds may be formulated into pharmaceutical compositions bycombination with appropriate, pharmaceutically acceptable carriers ordiluents, and may be formulated into preparations in solid, semi-solid,liquid or gaseous forms, such as tablets, capsules, powders, granules,ointments, solutions, suppositories, injections, inhalants and aerosols.In certain embodiments, the conjugate compounds are formulated forinjection. For example, compositions of interest may be formulated forintravenous or intraperitoneal administration.

In certain embodiments, compositions of interest include liposomal ormicellar compositions where the compounds described herein areliposome-based formulations or micelle-based formulations. Theliposome-based formulation or micelle-based formulation of the subjectcompounds may be prepared by any convenient liposome or micelle formingprotocol, such as for example by mechanical dispersion, solventdispersion, or a detergent removal method. In certain instances,liposomes are formed by mechanical dispersion including by sonication.French pressure cell extrusion, freeze-thawing, lipid film hydration(e.g., by hand-shaking, mechanical agitation or freeze drying),micro-emulsification, membrane extrusion or using dried reconstitutedvesicles. In certain embodiments, liposome-based formulations of thecompounds described herein are prepared by thin-film rehydrationfollowed by extrusion (e.g., through a filter of 5 nm or more, such as10 nm or more, such as 25 nm or more, such as 50 nm or more, such as 100nm or more, such as 150 nm or more, such as 200 nm or more, such as 250nm or more, such as 300 nm or more and including extrusion through afiler of 500 nm or more).

In some embodiments, the liposome-based formulation or micelle-basedformulation may be formed from a non-polymeric carrier materialincluding but not limited to: sterols such as cholesterol, stigmasterol,β-sitosterol, and estradiol; cholestery esters such as cholesterylstearate; C₁₂-C₂₄ fatty acids such as lauric acid, myristic acid,palmitic acid, stearic acid, arachidic acid, behenic acid, andlignoceric acid; C₁₈-C₃₆ mono-, di- and triacylglycerides such asglyceryl monooleate, glyceryl monolinoleate, glyceryl monolaurate,glyceryl monodocosanoate, glyceryl monomyristate, glycerylmonodicenoate, glyceryl dipalmitate, glyceryl didocosanoate, glyceryldimyristate, glyceryl didecenoate, glyceryl tridocosanoate, glyceryltrimyristate, glyceryl tridecenoate, glycerol tristearate and mixturesthereof; sucrose fatty acid esters such as sucrose distearate andsucrose palmitate; sorbitan fatty acid esters such as sorbitanmonostearate, sorbitan monopalmitate and sorbitan tristearate, C₁₆-C₁₈fatty alcohols such as cetyl alcohol, myristyl alcohol, stearyl alcohol,and cetostearyl alcohol; esters of fatty alcohols and fatty acids suchas cetyl palmitate and cetearyl palmitate; anhydrides of fatty acidssuch as stearic anhydride; phospholipids including phosphatidylcholine(lecithin), phosphatidylserine, phosphoethanolamine,phosphoethanolamine-PEG(2000), phosphatidylethanolamine,phosphatidylinositol, and lysoderivatives thereof; sphingosine andderivatives thereof; sphingomyelins such as stearyl, palmitoyl, andtricosanoyl sphingomyelins; ceramides such as stearyl and palmitoylceramides; glycosphingolipids; lanolin and lanolin alcohols; andcombinations and mixtures thereof. In certain embodiments,liposome-formulated compounds include phosphatidylcholine andcholesterol.

Each component used to prepare the liposome formulation or micelleformulation may vary as desired and may be present in an amount of 0.001wt % or more for the liposome or micelle formulation, such as 0.005 wt %or more, such as 0.010 wt % or more, such as 0.05 wt % or more, such as0.1 wt % or more, such as 0.5 wt % or more, such as 1 wt % or more, suchas 2 wt % or more, such as 3 wt % or more, such as 4 wt % or more andincluding where each component is present an in amount of 5 wt % ormore. Where more than one component is present (e.g., a phospholipidsuch as phosphatidylcholine and cholesterol), the ratio of thecomponents may range from 0.001:1 to 1:0.001, such as from 0.005:1 to1:0.005, such as from 0.01:1 to 1:0.01, such as from 0.05:1 to 1:0.05,such as from 0.1:1 to 1:0.1, such as from 0.5:1 to 1:0.5, such as from0.6:1 to 1:0.6, such as from 0.7:1 to 1:0.7, such as from 0.8:1 to1:0.8, such as from 0.9:1 to 1:0.9 and including where the ratio of thecomponents is 1:1 (e.g., phosphatidylcholine to cholesterol ratio of1:1). In one example, the liposome or micelle formulation may include aphospholipid component (e.g., phosphatidylcholine) and cholesterol, suchas in a ratio that ranges from 0.001:1 to 1:0.001, such as from 0.005:1to 1:0.005, such as from 0.01:1 to 1:0.01, such as from 0.05:1 to1:0.05, such as from 0.1:1 to 1:0.1, such as from 0.5:1 to 1:0.5, suchas from 0.6:1 to 1:0.6, such as from 0.7:1 to 1:0.7, such as from 0.8:1to 1:0.8, such as from 0.9:1 to 1:0.9 and including where the ratio ofthe components is 1:1 In certain instances, the ratio of phospholipidcomponent to cholesterol is about 2:1.

The compounds described herein (e.g., compounds of formula DCD-(I) orDCD-(II)) may be present in the liposome or micelle formulation in anamount of 0.001 wt % or more of the formulation, such as 0.005 wt % ormore, such as 0.010 wt % or more, such as 0.05 wt % or more, such as 0.1wt % or more, such as 0.5 wt % or more, such as 1 wt % or more, such as2 wt % or more, such as 3 wt % or more, such as 4 wt % or more andincluding where the active agent compound is present an in amount of 5wt % or more. The ratio of active agent compound (e.g., compound foractivating invariant natural killer T cells such as a compound offormula DCD-(I) of DCD-(II)) to each liposome component may range from0.001:1 to 1:0.001, such as from 0.005:1 to 1:0.005, such as from 0.01:1to 1:0.01, such as from 0.05:1 to 1:0.05, such as from 0.1:1 to 1:0.1,such as from 0.5:1 to 1:0.5, such as from 0.6:1 to 1:0.6, such as from0.7:1 to 1:0.7, such as from 0.8:1 to 1:0.8, such as from 0.9:1 to 1:0.9and including a ratio of the components is 1:1. In certain instances,the ratio is from 1:0.15 or 2:0.15. For example, a composition mayinclude phosphatidylcholine, cholesterol and an active agent compound ata ratio of 2:1:0.15.

In certain embodiments, liposome formulations or micelle formulationsmay include an organic solvent, such for example one or more organicsolvents selected from sterols such as cholesterol, stigmasterol,β-sitosterol, and estradiol; cholestery esters such as cholesterylstearate; C12-C24 fatty acids such as lauric acid, myristic acid,palmitic acid, stearic acid, arachidic acid, behenic acid, andlignoceric acid; C18-C36 mono-, di- and triacylglycerides such asglyceryl monooleate, glyceryl monolinoleate, glyceryl monolaurate,glyceryl monodocosanoate, glyceryl monomyristate, glycerylmonodicenoate, glyceryl dipalmitate, glyceryl didocosanoate, glyceryldimyristate, glyceryl didecenoate, glyceryl tridocosanoate, glyceryltrimyristate, glyceryl tridecenoate, glycerol tristearate and mixturesthereof; sucrose fatty acid esters such as sucrose distearate andsucrose palmitate; sorbitan fatty acid esters such as sorbitanmonostearate, sorbitan monopalmitate and sorbitan tristearate; C16-C18fatty alcohols such as cetyl alcohol, myristyl alcohol, stearyl alcohol,and cetostearyl alcohol; esters of fatty alcohols and fatty acids suchas cetyl palmitate and cetearyl palmitate; anhydrides of fatty acidssuch as stearic anhydride; phospholipids including phosphatidylcholine(lecithin), phosphatidylserine, phosphoethanolamine,phosphoethanolamine-PEG(2000), phosphatidylethanolamine,phosphatidylinositol, and lysoderivatives thereof, sphingosine andderivatives thereof; sphingomyelins such as stearyl, palmitoyl, andtricosanoyl sphingomyelins; ceramides such as stearyl and palmitoylceramides; glycosphingolipids; lanolin and lanolin alcohols; andcombinations and mixtures thereof.

In pharmaceutical dosage forms, the compounds may be administered in theform of its pharmaceutically acceptable salts, or it may also be usedalone or in appropriate association, as well as in combination, withother pharmaceutically active compounds. The following methods andexcipients are merely exemplary and are in no way limiting.

In some embodiments, compositions of interest include an aqueous buffer.Suitable aqueous buffers include, but are not limited to, acetate,succinate, citrate, and phosphate buffers varying in strengths fromabout 5 mM to about 100 mM. In some embodiments, the aqueous bufferincludes reagents that provide for an isotonic solution. Such reagentsinclude, but are not limited to, sodium chloride; and sugars e.g.,mannitol, dextrose, sucrose, and the like. In some embodiments, theaqueous buffer further includes a non-ionic surfactant such aspolysorbate 20 or 80. In some instances, compositions of interestfurther include a preservative. Suitable preservatives include, but arenot limited to, a benzyl alcohol, phenol, chlorobutanol, benzalkoniumchloride, and the like. In many cases, the composition is stored atabout 4° C. Formulations may also be lyophilized, in which case theygenerally include cryoprotectants such as sucrose, trehalose, lactose,maltose, mannitol, and the like. Lyophilized formulations can be storedover extended periods of time, even at ambient temperatures.

In some embodiments, compositions include other additives, such aslactose, mannitol, corn starch or potato starch; with binders, such ascrystalline cellulose, cellulose derivatives, acacia, corn starch orgelatins; with disintegrators, such as corn starch, potato starch orsodium carboxymethylcellulose; with lubricants, such as talc ormagnesium stearate; and if desired, with diluents, buffering agents,moistening agents, preservatives and flavoring agents.

Where the composition is formulated for injection, the compounds may beformulated by dissolving, suspending or emulsifying them in an aqueousor nonaqueous solvent, such as vegetable or other similar oils,synthetic aliphatic acid glycerides, esters of higher aliphatic acids orpropylene glycol; and if desired, with conventional additives such assolubilizers, isotonic agents, suspending agents, emulsifying agents,stabilizers and preservatives.

Although the dosage used in treating a subject will vary depending onthe clinical goals to be achieved, a suitable dosage range of thecompound is one which provides up to about 0.0001 mg to about 5000 mg,e.g., from about 1 mg to about 25 mg, from about 25 mg to about 50 mg,from about 50 mg to about 100 mg, from about 100 mg to about 200 mg,from about 200 mg to about 250 mg, from about 250 mg to about 500 mg,from about 500 mg to about 1000 mg, or from about 1000 mg to about 5000mg of an active agent, which can be administered in a single dose. Thoseof skill will readily appreciate that dose levels can vary as a functionof the specific compound, the severity of the symptoms and thesusceptibility of the subject to side effects.

In some embodiments, a single dose of the compound is administered. Inother embodiments, multiple doses of the compound are administered.Where multiple doses are administered over a period of time, thecompound may be administered, e.g., twice daily (qid), daily (qd), everyother day (qod), every third day, three times per week (tiw), or twiceper week (biw) over a period of time. For example, the compound may beadministered qid, qd, qod, tiw, or biw over a period of from one day toabout 2 years or more. For example, the compound may be administered atany of the aforementioned frequencies for one week, two weeks, onemonth, two months, six months, one year, or two years, or more,depending on various factors.

Dose units of the present disclosure can be made using manufacturingmethods available in the art and can be of a variety of forms suitablefor injection (including topical, intracisternal, intrathecal,intravenous, intramuscular, subcutaneous and dermal) administration, forexample as a solution, suspension, solution, lyophilate or emulsion. Thedose unit can contain components conventional in pharmaceuticalpreparations, e.g. one or more carriers, binders, lubricants, excipients(e.g., to impart controlled release characteristics), pH modifiers,coloring agents or further active agents.

Dose units can comprise components in any relative amounts. For example,dose units can be from about 0.1% to 99% by weight of active ingredients(i.e., compounds described herein) per total weight of dose unit. Insome embodiments, dose units can be from 10% to 50%, from 20% to 40%, orabout 30% by weight of active ingredients per total weight dose unit.

Aspects, including embodiments, of the subject matter described hereinmay be beneficial alone or in combination, with one or more otheraspects or embodiments. Without limiting the description, certainnon-limiting aspects of the disclosure numbered 1-99 are provided below.As will be apparent to those of skill in the art upon reading thisdisclosure, each of the individually numbered aspects may be used orcombined with any of the preceding or following individually numberedaspects. This is intended to provide support for all such combinationsof aspects and is not limited to combinations of aspects explicitlyprovided below:

-   -   1. A compound of formula DCD-(I):

-   -   wherein:    -   Z is selected from:

-   -   wherein        indicates the Z—O bond;    -   wherein X is —NHCO— or oxygen;    -   R¹, R², R³ and R⁴ are each independently selected from hydrogen,        alkyl, substituted alkyl, heteroalkyl, substituted heteroalkyl,        cycloalkyl, substituted cycloalkyl, heterocycloalkyl,        substituted heterocycloalkyl, aryl, substituted aryl, arylalkyl,        substituted arylalkyl, heteroaryl, substituted heteroaryl,        heteroarylalkyl, and substituted heteroarylalkyl;    -   X₁ and X₂ are each independently selected from —C, —NR^(j), —O,        —SR^(k), —Si, wherein R^(j) and R^(k) are each independently        selected from hydrogen, alkyl or substituted alkyl, heteroalkyl,        substituted heteroalkyl, cycloalkyl, substituted cycloalkyl,        heterocycloalkyl, substituted heterocycloalkyl, aryl,        substituted aryl, arylalkyl, substituted arylalkyl, heteroaryl,        substituted heteroaryl, heteroarylalkyl, and substituted        heteroarylalkyl;    -   R^(a) is selected from hydrogen, oxygen, fluorine, —CF₃, or        together with X₂ form a cycloalkyl, substituted cycloalkyl,        heterocycloalkyl, substituted heterocycloalkyl, aryl,        substituted aryl, arylalkyl, substituted arylalkyl, heteroaryl,        substituted heteroaryl, heteroarylalkyl, and substituted        heteroarylalkyl;    -   wherein        indicates a double or single bond;    -   n is an integer from 2 to 25,    -   Y is selected from carbon, nitrogen or silicon;    -   R^(b), R^(c) and R^(d) are independently selected hydrogen,        alkyl, substituted alkyl, heteroalkyl, substituted heteroalkyl,        cycloalkyl, substituted cycloalkyl, heterocycloalkyl,        substituted heterocycloalkyl, aryl, substituted aryl, arylalkyl,        substituted arylalkyl, heteroaryl, substituted heteroaryl,        heteroarylalkyl, and substituted heteroarylalkyl, wherein when Y        is nitrogen, R^(d) is not present, or wherein R^(c) and R^(d)        together with Y form a cycloalkyl, substituted cycloalkyl, aryl,        substituted aryl, heteroaryl or substituted heteroaryl group;        and    -   R^(e) is alkyl or substituted alkyl,    -   or salt, solvate or hydrate thereof.    -   2. The compound according to 1, wherein R¹, R², R³ and R⁴ are        each hydrogen.    -   3. The compound according to 1, wherein R¹ is hydrogen.    -   4. The compound according to any one of 1-3, wherein R¹ is:

-   -   wherein        indicates the R¹—O bond;    -   R⁸ is hydrogen, alkyl, substitute alkyl;    -   R⁹ is —NR^(f) or —OR^(f), wherein R^(f) is alkyl, substituted        alkyl, acyl, alkylacyl or substituted alkylacyl, or wherein        R^(f) together with R¹⁰ form a cycloalkyl, substituted        cycloalkyl, heterocycloalkyl, substituted heterocycloalkyl,        aryl, substituted aryl, arylalkyl, substituted arylalkyl,        heteroaryl, substituted heteroaryl, heteroarylalkyl, and        substituted heteroarylalkyl; and    -   R¹⁰ is cycloalkyl, substituted cycloalkyl, heterocycloalkyl,        substituted heterocycloalkyl, aryl, substituted aryl, arylalkyl,        substituted arylalkyl, heteroaryl, substituted heteroaryl,        heteroarylalkyl, and substituted heteroarylalkyl or wherein R¹⁰        together with R^(f) form a cycloalkyl, substituted cycloalkyl,        heterocycloalkyl, substituted heterocycloalkyl, aryl,        substituted aryl, arylalkyl, substituted arylalkyl, heteroaryl,        substituted heteroaryl, heteroarylalkyl, and substituted        heteroarylalkyl.    -   5. The compound according to 4, wherein R¹ is selected from:

-   -   wherein        indicates the R¹—O bond;    -   R⁸ is hydrogen, alkyl, substitute alkyl; and    -   R^(g) is hydrogen or a halogen selected from fluorine, chlorine,        bromine or iodine.    -   6. The compound according to any one of 1-3, wherein R¹ is:

-   -   wherein:    -   R¹¹ is alkyl or substituted alkyl;    -   R¹² is alkyl or alkyl substituted with a cycloalkyl, substituted        cycloalkyl, heterocycloalkyl, substituted heterocycloalkyl,        aryl, substituted aryl, arylalkyl, substituted arylalkyl,        heteroaryl, substituted heteroaryl, heteroarylalkyl, and        substituted heteroarylalkyl.    -   7. The compound according to 6, wherein R¹ is:

-   -   wherein R^(h) and R^(i) are each independently selected from        hydrogen, hydroxyl or a halogen selected from F, Cl, I or Br.    -   8. The compound according to any one of 1-7, wherein X₁ is —NH.    -   9. The compound according to 8, wherein:        -   R^(a) is O;        -   X₂ is C; and

is carbonyl.

-   -   10. The compound according to 8, wherein:        -   R^(a) is O;        -   X₂ is —SR^(k), wherein R^(k) is methyl; and

is sulfur oxide.

-   -   11. The compound according to 8, wherein:        -   R^(a) is CF₃; and        -   X₂ is C.    -   12. The compound according to 8, wherein R^(a) together with X²        form a heterocycloalkyl or substituted heterocycloalkyl.    -   13. The compound according to 12, wherein R^(a) together with X₂        forms an oxacyclobutane.    -   14. The compound according to any one of 1-7, wherein:        -   R^(a) is F;        -   X₁ is C;        -   X₂ is C; and        -   is a double bond.    -   15. The compound according to any one of 1-14, wherein R^(e) is        a C8 to C20 alkyl or substituted C8 to C20 alkyl.    -   16. The compound according to 19, wherein R^(e) is a C13 alkyl.    -   17. The compound according to any one of 1-16, wherein R^(d) is        a C5 to C25 alkyl or a C5 to C25 alkyl substituted with a        cycloalkyl group, substituted cycloalkyl group, heterocycloalkyl        group, substituted heterocycloalkyl group, aryl group,        substituted aryl group, arylalkyl group, substituted arylalkyl        group, heteroaryl group, substituted heteroaryl group,        heteroarylalkyl group, or substituted heteroarylalkyl group.    -   18. The compound according to 17, wherein R^(d) is a C5 to C25        alkyl substituted with a moiety selected from the group        consisting of:

-   -   wherein        indicates the bond to the C5 to C25 alkyl;    -   R^(m) and R^(n) are independently selected from hydrogen,        halogen, hydroxyl, substituted hydroxyl, amino, substituted        amino, thiol, substituted thiol, sulfoxide, substituted        sulfoxide, sulfone, substituted sulfone, sulfoximine,        substituted sulfoximine, acyl, aminoacyl, alkyl, substituted        alkyl; heteroalkyl, substituted heteroalkyl, cycloalkyl,        substituted cycloalkyl, spiroalkyl, heterocycloalkyl,        substituted heterocycloalkyl, aryl, substituted aryl, arylalkyl,        substituted arylalkyl, heteroaryl, substituted heteroaryl,        heteroarylalkyl, and substituted heteroarylalkyl.    -   19. The compound according to 18, wherein R^(m) is hydrogen.    -   20. The compound according to 18, wherein R^(m) is halogen.    -   21. The compound according to 20, wherein R^(m) is selected from        fluorine, bromine or iodine.    -   22. The compound according to any one of 18-21, wherein R^(n) is        hydrogen.    -   23. The compound according to 18-21, wherein R^(n) is halogen.    -   24. The compound according to 23, wherein R^(n) is fluorine,        bromine or iodine.    -   25. The compound according to any one of 1-24, wherein R^(b) is        hydrogen.    -   26. The compound according to any one of 1-24, wherein R^(b) is        selected from the group consisting of methyl, ethyl, propyl,        butyl and

-   -   wherein        indicates a bond to Y and R^(z) is hydrogen, alkyl or alkyl        substituted with a cycloalkyl, substituted cycloalkyl,        heterocycloalkyl, substituted heterocycloalkyl, aryl,        substituted aryl, arylalkyl, substituted arylalkyl, heteroaryl,        substituted heteroaryl, heteroarylalkyl, and substituted        heteroarylalkyl. In certain instances, R is alkyl, such as a C1        to C16 alkyl or C1 to C16 substituted alkyl.    -   27. The compound according to any one of 1-24, wherein R^(b) is:

wherein

indicates a bond to Y and R^(z) is hydrogen, alkyl or alkyl substitutedwith a cycloalkyl, substituted cycloalkyl, heterocycloalkyl, substitutedheterocycloalkyl, aryl, substituted aryl, arylalkyl, substitutedarylalkyl, heteroaryl, substituted heteroaryl, heteroarylalkyl, andsubstituted heteroarylalkyl. In certain instances, R^(z) is alkyl, suchas a C1 to C16 alkyl or C1 to C16 substituted alkyl.

-   -   28. The compound according to any one of 1-24, wherein R^(b) is:

wherein

indicates a bond to Y and R^(z) is hydrogen, alkyl or alkyl substitutedwith a cycloalkyl, substituted cycloalkyl, heterocycloalkyl, substitutedheterocycloalkyl, aryl, substituted aryl, arylalkyl, substitutedarylalkyl, heteroaryl, substituted heteroaryl, heteroarylalkyl, andsubstituted heteroarylalkyl. In certain instances, R^(z) is alkyl, suchas a C1 to C16 alkyl or C1 to C16 substituted alkyl.

-   -   29. The compound according to any one of 1-28, wherein R^(c) is        a C1 to C10 alkyl.    -   30. The compound according to 26, wherein R^(c) is selected from        the group consisting of methyl, ethyl, propyl and butyl.    -   31. The compound according to 1, wherein the compound is        selected from the group consisting of:

Compound No. Structure DCD-101

DCD-113

DCD-102

DCD-103

DCD-106

-   -   32. A method comprising contacting invariant natural killer T        (iNKT) cells with a compound according to any one of 1-31.    -   33. The method according to 32, wherein the iNKT cells are        contacted to in vitro.    -   34. The method according to 32, wherein the iNKT cells are        contacted in vivo.    -   35. The method according to any one of 32-34, wherein contacting        the compound with the iNKT cells is sufficient to activate the        iNKT cells.    -   36. The method according to 35, wherein activating the iNKT        cells induce an increase in production of one or more cytokines        selected from the group consisting of IFN-γ, IL-1β, IL-2, IL-3,        IL-8, IL-12, IL-15, TNF-α, GM-CSF, RANTES, MIP-1α and MCP-1.    -   37. The method according to 35, wherein activating the iNKT        cells induce an increase in production of one or more cytokines        selected from the group consisting of IL-4, IL-6, IL-8, IL-10        and IL-13.    -   38. The method according to any one of 35-37, wherein the method        further comprises contacting the activated iNKT cells with a        composition comprising senescent cells, wherein contacting the        activated iNKT cells reduces the presence of or eliminates the        senescent cells in the composition.    -   39. The method according to 39, wherein the senescent cells        comprise an inflammatory secretome.    -   40. The method according to any one of 38-39, wherein the        composition further comprises healthy cells.    -   41. The method according to 40, wherein contacting the activated        iNKT cells reduces the presence of or eliminates the senescent        cells in the composition without reducing the presence of the        healthy cells.    -   42. The method according to 41, wherein the presence of healthy        cells is reduced by 5% or less when the composition is contacted        with the activated iNKT cells.    -   43. A method comprising administering a compound according to        any one of 1-31 to a subject in need thereof.    -   44. A method for selectively reducing the presence of or        eliminating senescent cells in a subject, the method comprising        administering a compound according to any one of 1-31 to a        subject in need thereof.    -   45. The method according to 36, wherein the subject is diagnosed        as having an autoimmune disease, an allergic disease, a        metabolic disorder, cancer or a pathogen infection.    -   46. The method according to 45, wherein the subject is diagnosed        as having one or more of a metabolic disorder, an eye disease, a        disease of aging, fibrosis, heart disease, kidney disease.    -   47. A pharmaceutical composition comprising:        -   a compound according to any one of 1-31; and        -   a pharmaceutically acceptable carrier.    -   48. A pharmaceutical composition for selectively reducing the        presence of or eliminating senescent cells in a subject, the        composition comprising:        -   a compound according to any one of 1-31; and        -   a pharmaceutically acceptable carrier.    -   49. Use of a compound according to any one of 1-31 in the        manufacture of a medicament for treating a subject in need        thereof.    -   50. Use of a compound according to any one of 1-31 in the        manufacture of a medicament for selectively reducing the        presence of or eliminating senescent cells in a subject in need        thereof.    -   51. A compound of formula DCD-(11):

-   -   wherein:    -   Z is selected from:

-   -   wherein        indicates the Z—O bond;    -   wherein X is —NHCO— or oxygen;    -   R¹, R², R³ and R⁴ are each independently selected from hydrogen,        alkyl, substituted alkyl, heteroalkyl, substituted heteroalkyl,        cycloalkyl, substituted cycloalkyl, heterocycloalkyl,        substituted heterocycloalkyl, aryl, substituted aryl, arylalkyl,        substituted aryalkyl, heteroaryl, substituted heteroaryl,        heteroarylalkyl, and substituted heteroarylalkyl;    -   X₁, X₂, X₃, X₄ and X₅ are each independently selected from        carbon, nitrogen, oxygen or sulfur;    -   R^(a) is optionally absent or when present is selected from        hydrogen or oxygen;    -   wherein        indicates a double or single bond,    -   n is an integer from 2 to 25;    -   Y is selected from carbon, nitrogen or silicon;    -   R^(b), R^(c) and R^(d) are independently selected hydrogen,        alkyl, substituted alkyl, heteroalkyl, substituted heteroalkyl,        cycloalkyl, substituted cycloalkyl, heterocycloalkyl,        substituted heterocycloalkyl, aryl, substituted aryl, arylalkyl,        substituted arylalkyl, heteroaryl, substituted heteroaryl,        heteroarylalkyl, and substituted heteroarylalkyl, wherein when Y        is nitrogen, R^(d) is not present, or wherein R^(c) and R^(d)        together with Y form a cycloalkyl, substituted cycloalkyl, aryl,        substituted aryl, heteroaryl or substituted heteroaryl group;        and    -   R^(e) is alkyl or substituted alkyl, or salt, solvate or hydrate        thereof.    -   52. The compound according to 51, wherein R¹, R², R³ and R⁴ are        each hydrogen.    -   53. The compound according to 52, wherein R¹ is hydrogen.    -   54. The compound according to any one of 51-53, wherein R¹ is:

-   -   wherein        indicates the R¹—O bond;    -   R^(d) is hydrogen, alkyl, substitute alkyl;    -   R⁹ is —NR^(f) or —OR^(f),        -   wherein R^(f) is alkyl, substituted alkyl, acyl, alkylacyl            or substituted alkylacyl,        -   or wherein R^(f) together with R¹⁰ form a cycloalkyl,            substituted cycloalkyl, heterocycloalkyl, substituted            heterocycloalkyl, aryl, substituted aryl, arylalkyl,            substituted arylalkyl, heteroaryl, substituted heteroaryl,            heteroarylalkyl, and substituted heteroarylalkyl; and    -   R¹⁰ is cycloalkyl, substituted cycloalkyl, heterocycloalkyl,        substituted heterocycloalkyl, aryl, substituted aryl, arylalkyl,        substituted arylalkyl, heteroaryl, substituted heteroaryl,        heteroarylalkyl, and substituted heteroarylalkyl or wherein R¹⁰        together with R^(f) form a cycloalkyl, substituted cycloalkyl,        heterocycloalkyl, substituted heterocycloalkyl, aryl,        substituted aryl, arylalkyl, substituted arylalkyl, heteroaryl,        substituted heteroaryl, heteroarylalkyl, and substituted        heteroarylalkyl.    -   55. The compound according to 54, wherein R¹ is selected from:

-   -   wherein        indicates the R¹—O bond;    -   R⁸ is hydrogen, alkyl, substitute alkyl; and    -   R^(g) is hydrogen or a halogen selected from F, Cl, I or Br.    -   56. The compound according to any one of 51-55, wherein R¹ is:

-   -   wherein:    -   R¹¹ is alkyl or substituted alkyl;    -   R¹² is alkyl or alkyl substituted with a cycloalkyl, substituted        cycloalkyl, heterocycloalkyl, substituted heterocycloalkyl,        aryl, substituted aryl, arylalkyl, substituted arylalkyl,        heteroaryl, substituted heteroaryl, heteroarylalkyl, and        substituted heteroarylalkyl.    -   57. The compound according to 56, wherein R¹ is:

-   -   wherein R^(h) and R^(i) are each independently selected from        hydrogen, hydroxyl or a halogen selected from F, Cl, I or Br.    -   58. The compound according to any one of 51-57, wherein X₁, X₂,        X₃, X₄ and X₅ together form a pyrazole.    -   59. The compound according to any one of 51-57, wherein X₁, X₂,        X₃, X₄ and X₅ together form an imidazole.    -   60. The compound according to any one of 51-57, wherein X₁, X₂,        X₃, X₄ and X₅ together form a tetrazole.    -   61. The compound according to any one of 51-57, wherein:        -   X₁ is carbon;        -   X₂ is nitrogen;        -   X₃ is nitrogen;        -   X₄ is carbon; and        -   X₅ is carbon.    -   62. The compound according to any one of 51-57, wherein:        -   X₁ is carbon;        -   X₂ is nitrogen;        -   X₃ is carbon;        -   X₄ is carbon; and        -   X₅ is nitrogen.    -   63. The compound according to any one of 51-57, wherein:        -   X₁ is nitrogen;        -   X₂ is nitrogen;        -   X₃ is nitrogen;        -   X₄ is nitrogen;        -   X₅ is carbon;        -   R^(a) is O,    -   wherein

is carbonyl.

-   -   64. The compound according to any one of 51-63, wherein R^(e) is        a C8 to C20 alkyl or substituted C8 to C20 alkyl.    -   65. The compound according to 64, wherein R^(e) is a C13 alkyl.    -   66. The compound according to any one of 51-65, wherein R^(d) is        a C5 to C25 alkyl or a C5 to C25 alkyl substituted with a        cycloalkyl group, substituted cycloalkyl group, heterocycloalkyl        group, substituted heterocycloalkyl group, aryl group,        substituted aryl group, arylalkyl group, substituted arylalkyl        group, heteroaryl group, substituted heteroaryl group,        heteroarylalkyl group, or substituted heteroarylalkyl group    -   67, The compound according to 66, wherein R^(d) is a C5 to C25        alkyl substituted with a moiety selected from the group        consisting of:

-   -   wherein        indicates the bond to the C5 to C25 alkyl; and    -   R^(m) and R^(n) are independently selected from hydrogen,        halogen, hydroxyl, substituted hydroxyl, amino, substituted        amino, thiol, substituted thiol, sulfoxide, substituted        sulfoxide, sulfone, substituted sulfone, sulfoximine,        substituted sulfoximine, acyl, aminoacyl, alkyl, substituted        alkyl; heteroalkyl, substituted heteroalkyl, cycloalkyl,        substituted cycloalkyl, spiroalkyl, heterocycloalkyl,        substituted heterocycloalkyl, aryl, substituted aryl, arylalkyl,        substituted arylalkyl, heteroaryl, substituted heteroaryl,        heteroarylalkyl, and substituted heteroarylalkyl.    -   68. The compound according to 67, wherein R^(m) is hydrogen.    -   69. The compound according to 67, wherein R^(m) is halogen.    -   70. The compound according to 69, wherein R^(m) is selected from        fluorine, bromine or iodine.    -   71. The compound according to any one of 67-70, wherein R^(n) is        hydrogen.    -   72. The compound according to 67-70, wherein R^(n) is halogen.    -   73. The compound according to 72, wherein R^(n) is fluorine,        bromine or iodine.    -   74. The compound according to any one of 51-73, wherein R^(b) is        hydrogen.    -   75. The compound according to any one of 51-73, wherein R^(b) is        selected from the group consisting of methyl, ethyl, propyl,        butyl and

-   -   wherein        indicates a bond to Y and R^(z) is hydrogen, alkyl or alkyl        substituted with a cycloalkyl, substituted cycloalkyl,        heterocycloalkyl, substituted heterocycloalkyl, aryl,        substituted aryl, arylalkyl, substituted arylalkyl, heteroaryl,        substituted heteroaryl, heteroarylalkyl, and substituted        heteroarylalkyl. In certain instances, R² is alkyl, such as a C1        to C16 alkyl or C1 to C16 substituted alkyl.    -   76. The compound according to any one of 51-73, wherein R^(b)        is:

wherein

indicates a bond to Y and R^(z) is hydrogen, alkyl or alkyl substitutedwith a cycloalkyl, substituted cycloalkyl, heterocycloalkyl, substitutedheterocycloalkyl, aryl, substituted aryl, arylalkyl, substitutedarylalkyl, heteroaryl, substituted heteroaryl, heteroarylalkyl, andsubstituted heteroarylalkyl. In certain instances, R² is alkyl, such asa C1 to C16 alkyl or C1 to C16 substituted alkyl.

-   -   77. The compound according to any one of 51-73, wherein R^(b)        is:

wherein

indicates a bond to Y and R^(z) is hydrogen, alkyl or alkyl substitutedwith a cycloalkyl, substituted cycloalkyl, heterocycloalkyl, substitutedheterocycloalkyl, aryl, substituted aryl, arylalkyl, substitutedarylalkyl, heteroaryl, substituted heteroaryl, heteroarylalkyl, andsubstituted heteroarylalkyl. In certain instances, R^(z) is alkyl, suchas a C1 to C16 alkyl or C1 to C16 substituted alkyl.

-   -   78. The compound according to any one of 51-77, wherein R^(c) is        a C1 to C10 alkyl.    -   79. The compound according to 78, wherein R^(c) is selected from        the group consisting of methyl, ethyl, propyl and butyl.    -   80. The compound according to 51, wherein the compound is        selected from Compound DCD-104 and DCD-105:

-   -   81. A method comprising contacting invariant natural killer T        (iNKT) cells with a compound according to any one of 51-80.    -   82. The method according to 81, wherein the iNKT cells are        contacted in vitro.    -   83. The method according to 81, wherein the iNKT cells are        contacted in vivo.    -   84. The method according to any one of 81-83, wherein contacting        the compound with the iNKT cells is sufficient to activate the        iNKT cells.    -   85. The method according to 84, wherein activating the iNKT        cells induce an increase in production of one or more cytokines        selected from the group consisting of IFN-γ, IL-1β, IL-2, IL-3,        IL-8, IL-12, IL-15, TNF-α, GM-CSF, RANTES, MIP-1α and MCP-1.    -   86. The method according to 84, wherein activating the iNKT        cells induce an increase in production of one or more cytokines        selected from the group consisting of IL-4, IL-6, IL-8, IL-10        and IL-13.    -   87. The method according to any one of 84-86, wherein the method        further comprises contacting the activated iNKT cells with a        composition comprising senescent cells, wherein contacting the        activated iNKT cells reduces the presence of or eliminates the        senescent cells in the composition.    -   88. The method according to 87, wherein the senescent cells        comprise an inflammatory secretome.    -   89. The method according to any one of 87-88, wherein the        composition further comprises healthy cells.    -   90. The method according to 89, wherein contacting the activated        iNKT cells reduces the presence of or eliminates the senescent        cells in the composition without reducing the presence of the        healthy cells.    -   91. The method according to 90, wherein the presence of healthy        cells is reduced by 5% or less when the composition is contacted        with the activated iNKT cells.    -   92. A method comprising administering a compound according to        any one of 51-80 to a subject in need thereof.    -   93. A method for selectively reducing the presence of or        eliminating senescent cells in a subject, the method comprising        administering a compound according to any one of 51-80 to a        subject in need thereof.    -   94. The method according to 93, wherein the subject is diagnosed        as having an autoimmune disease, an allergic disease, a        metabolic disorder, cancer or a pathogen infection.    -   95. The method according to 93, wherein the subject is diagnosed        as having one or more of a metabolic disorder, an eye disease, a        disease of aging, fibrosis, heart disease, kidney disease.    -   96. A pharmaceutical composition comprising:        -   a compound according to any one of 51-80; and        -   a pharmaceutically acceptable carrier.    -   97. A pharmaceutical composition for selectively reducing the        presence of or eliminating senescent cells in a subject, the        composition comprising:        -   a compound according to any one of 51-80; and        -   a pharmaceutically acceptable carrier.    -   98. Use of a compound according to any one of 51-80 in the        manufacture of a medicament for treating a subject in need        thereof.    -   99. Use of a compound according to any one of 51-80 in the        manufacture of a medicament for selectively reducing the        presence of or eliminating senescent cells in a subject in need        thereof.

EXAMPLES

The following examples are put forth so as to provide those of ordinaryskill in the art with a complete disclosure and description of how tomake and use the present invention, and are not intended to limit thescope of what the inventors regard as their invention nor are theyintended to represent that the experiments below are all or the onlyexperiments performed. Efforts have been made to ensure accuracy withrespect to numbers used (e.g., amounts, temperature, etc.) but someexperimental errors and deviations should be accounted for.

Example 1—Synthesis of Compounds General Synthetic Procedures

Many general references providing commonly known chemical syntheticschemes and conditions useful for synthesizing the disclosed compoundsare available (see. e.g., Smith and March, March's Advanced OrganicChemistry: Reactions. Mechanisms, and Structure, Fifth Edition,Wiley-Interscience, 2001; or Vogel, A Textbook of Practical OrganicChemistry. Including Qualitative Organic Analysis. Fourth Edition, NewYork: Longman, 1978).

Compounds as described herein can be purified by any of the means knownin the art, including chromatographic means, such as high performanceliquid chromatography (HPLC), preparative thin layer chromatography,flash column chromatography and ion exchange chromatography. Anysuitable stationary phase can be used, including normal and reversedphases as well as ionic resins. See, e.g., Introduction to Modern LiquidChromatography, 2nd Edition, ed. L. R. Snyder and J. J. Kirkland, JohnWiley and Sons, 1979, and Thin Layer Chromatography, ed E. Stahl,Springer-Verlag, New York, 1969.

During any of the processes for preparation of the compounds of thepresent disclosure, it may be necessary and/or desirable to protectsensitive or reactive groups on any of the molecules concerned. This canbe achieved by means of conventional protecting groups as described instandard works, such as T. W. Greene and P. G. M. Wuts, “ProtectiveGroups in Organic Synthesis”. Fourth edition, Wiley, New York 2006. Theprotecting groups can be removed at a convenient subsequent stage usingmethods known from the art.

The compounds described herein can contain one or more chiral centersand/or double bonds and therefore, can exist as stereoisomers, such asdouble-bond isomers (i.e., geometric isomers), enantiomers ordiastereomers. Accordingly, all possible enantiomers and stereoisomersof the compounds including the stereoisomerically pure form (e.g.,geometrically pure, enantiomerically pure or diastereomerically pure)and enantiomeric and stereoisomeric mixtures are included in thedescription of the compounds herein. Enantiomeric and stereoisomericmixtures can be resolved into their component enantiomers orstereoisomers using separation techniques or chiral synthesis techniqueswell known to the skilled artisan. The compounds can also exist inseveral tautomeric forms including the enol form, the keto form andmixtures thereof. Accordingly, the chemical structures depicted hereinencompass all possible tautomeric forms of the illustrated compounds.The compounds described also include isotopically labeled compoundswhere one or more atoms have an atomic mass different from the atomicmass conventionally found in nature. Examples of isotopes that can beincorporated into the compounds disclosed herein include, but are notlimited to, ²H, ³H, ¹¹C, ¹³C, ¹⁴C, ¹⁵N, ¹⁸O, ¹⁷O, etc. Compounds canexist in unsolvated forms as well as solvated forms, including hydratedforms. In general, compounds can be hydrated or solvated. Certaincompounds can exist in multiple crystalline or amorphous forms. Ingeneral, all physical forms are equivalent for the uses contemplatedherein and are intended to be within the scope of the presentdisclosure.

The nomenclature used herein to name the subject compounds isillustrated in the Examples herein. When possible, this nomenclature hasgenerally been derived using the commercially-available Auto Nomsoftware (MDL, San Leandro. Calif.).

The compounds of the present disclosure may be prepared by variousmethods. Enclosed herein are exemplary methods of making compoundsdescribed herein.

Synthesis of (2S,3S,4R)-2-azidooctadecane-1,3,4-triol

Reference is made to ChemBioChem 2012, 13, 1689-1697 and Eur. J. Org.Chem, 1998, 291-229

A mixture of DCM (25 mL) and H₂O (25 mL) containing NaN₃ (10 g, 153mmol) cooled to 0° C., and Tf₂O (5.5 mL, 31.5 mmol) was added dropwiseover 20 min. After addition, the resulting mixture was stirred at rt for3 h. The organic layer was separated, and the aqueous portion wasextracted with DCM (2×50 mL). The combined organic layers were washedwith saturated aqueous Na₂CO₃ and used directly in the next step.

To a suspension of (2S,3S,4R)-2-aminooctadecane-1,3,4-triol (5 g, 15.5mmol), K₂CO₃ (10.9 g, 79.0 mmol), and CuSO₄ (100 mg) in a mixture ofMeOH (30 mL) and H₂O (30 mL) was added the above organic DCM layer,which contained TfN₃. More MeOH was added to make the mixture ahomogeneous solution. The reaction mixture was stirred overnight at roomtemperature. The organic solvent was removed under vacuum and theaqueous layer was extracted with EtOAc. The organic layer was dried overanhydrous Na₂SO₄, filtered and concentrated wider reduced pressure toafford a crude residue that was purified by silica gel columnchromatography (PE/EtOAc 1:1) to give(2S,3S,4R)-2-azidooctadecane-1,3,4-triol (4.5 g, 83%) as an oil.

Synthesis of (2S,3S,4R)-2-azido-1-(trityloxy)octadecane-3,4-diol

A mixture of (2S,3S,4R)-2-azidooctadecane-1,3,4-triol (6.00 g, 17.5mmol, 1.0 eq), TrtCl (6.8 g, 24.4 mmol, 1.4 eq) and DMAP (213 mg, 1.74mmol, 0.1 eq) in dry pyridine (100 mL) was stirred at 50° C. overnight.Pyridine was removed under reduced pressure and the residue was dilutedwith EtOAc (200 mL), washed with water (2×50 mL), brine (2×50 mL), driedover Na₂SO₄ and concentrated under reduced pressure. The residue waspurified by column chromatography on silica gel (120 g; EtOAc/PE 1:2) togive (2S,3S,4R)-2-azido-1-(trityloxy)octadecane-3,4-diol (9 g, 88%) asan oil.

Synthesis of(((2S,3S,4R)-2-azido-3,4-bis(benzyloxy)octadecyloxy)methanetriyl)tribenzene

To a solution of (2S,3S,4R)-2-azido-1-(trityloxy)octadecane-3,4-diol(9.0 g, 15.4 mmol, 1.0 eq) in dry DMF (120 mL) at 0° C. was added NaH,60% dispersion in oil (2.2 g, 55.0 mmol, 3.5 eq) in portions. Aftercomplete addition, the mixture was stirred at 0° C. for 10 min. BnBr(9.2 g, 53.8 mmol, 3.5 eq) was added and the mixture was allowed to warmto rt and stirred for an additional 5 h. It was poured into ice/water(200 mL), diluted with EtOAc (1 L), and the organic layer washed withwater (4×200 mL), brine (2×200 mL), dried over Na₂SO₄ and concentratedunder reduced pressure. The residue was purified by columnchromatography on silica gel (120 g; EtOAc/PE 1:15) to give(((2S,3S,4R)-2-azido-3,4-bis(benzyloxy)octadecyloxy)methanetriyl)tribenzene(10 g, 85%) as an oil. LC/MS: mass calcd. for C₅₁H₆₃N₃O₃: 765.49, found:788.50 [M+Na]⁺.

Synthesis of (2S,3S,4R)-2-azido-3,4-bis(benzyloxy)octadecan-1-ol

To a mixture of(((2S,3S,4R)-2-azido-3,4-bis(benzyloxy)octadecyloxy)methanetriyl)tribenzene(10 g, 13.1 mmol, 1.0 eq) in toluene (60 mL) and MeOH (60 mL) was addedconcentrated aqueous HCl (2 mL; 12 M). The mixture was heated to 60° C.,and stirred overnight. The pH value of the aqueous phase was adjusted to˜7 used 1 M NaOH and the mixture was concentrated under reducedpressure. The crude residue was purified by column chromatography onsilica gel (120 g; EtOAc/PE: 1:10) to give(2S,3S,4R)-2-azido-3,4-bis(benzyloxy)octadecan-1-ol (5 g, 73%) as anoil. LC/MS: mass calcd. for C₃₂H₄₄N₃O₃: 523.38, found: 546.25 [M+Na]⁺.

Synthesis of(3R,4S,5S,6R)-3,4,5-tris(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-2-ylAcetate

A mixture of(3R,4S,5S,6R)-3,4,5-tris(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-2-ol(25 g, 9.24 mmol), pyridine (60 mL) and Ac₂O (120 mL) was stirred at rtovernight. The reaction was quenched with crushed ice and the resultingmixture was extracted with DCM (3×300 mL). The combined organic layerswere concentrated under reduced pressure and the crude residue waspurified by column chromatography on silica gel (PE/EtOAc 1:1) to give(3R,4S,5S,6R)-3,4,5-tris(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-2-ylacetate (20 g, 74%) as an oil.

Synthesis of(2R,3S,4S,5R,6R)-3,4,5-tris(benzyloxy)-2-(benzyloxymethyl)-6-iodo-tetrahydro-2H-pyran

To a mixture of(3R,4S,5S,6R)-3,4,5-tris(benzyloxy)-6-(benzyloxymethyl)-tetrahydro-2H-pyran-2-ylacetate (20.0 g, 34.3 mmol, 1.0 eq) in DCM (150 mL) under an atmosphereof N₂ at 0° C. was added TMSI (6.9 g, 34.5 mmol, 1.0 eq). The mixturewas stirred at 0° C. for 40 min, then benzene (50 mL) added and themixture concentrated under reduced pressure to give(2R,3S,4S,5R,6R)-3,4,5-tris(benzyloxy)-2-(benzyloxymethyl)-6-iodo-tetrahydro-2H-pyran(19 g, 85%) of as an oil.

Synthesis of(2S,3R,4S,5S,6R)-2-((2S,3S,4R)-2-azido-3,4-bis(benzyloxy)octadecyloxy)-3,4,5-tris(benzyloxy)-6-(benzyloxymethyl)-tetrahydro-2H-pyran

A mixture of (2S,3S,4R)-2-azido-3,4-bis(benzyloxy)octadecan-1-ol (3.0 g,5.7 mmol, 1.0 eq), TBAI (19.0 g, 51.4 mmol, 9.0 eq), DIPEA (2.2 g, 17.0mmol, 3.0 eq) and 4 Å-MS (2 g) in benzene (80 mL) was stirred at 65° C.for 20 min under an atmosphere of N₂. To this mixture was added asolution of(2R,3S,4S,5R,6R)-3,4,5-tris(benzyloxy)-2-(benzyloxymethyl)-6-iodo-tetrahydro-2H-pyran(18.6 g, 17.2 mmol, 3.0 eq) in benzene (30 mL). The mixture was stirredat 65° C. for an additional 2 h, cooled rt and EtOAc (150 mL) added. Themixture was filtered, the filtrate was washed with saturated sodiumthiosulfate solution (2×80 mL), brine (2×80 mL), dried over Na₂SO₄ andconcentrated under reduced pressure. The residue was purified by columnchromatography on silica gel column (120 g; EtOAc/PE 1:5) to give(2S,3R,4S,5S,6R)-2-((2S,3S,4R)-2-azido-3,4-bis(benzyloxy)octadecyloxy)-3,4,5-tris(benzyloxy)-6-(benzyloxymethyl)-tetrahydro-2H-pyran(3 g, 50%) as an oil. LC/MS: mass calcd. for C₆₆H₈₃N₃O₈: 1045.62, found:1068.60 [M+Na]⁺.

Synthesis of(2S,3S,4R)-3,4-bis(benzyloxy)-1-((2S,3R,4S,5S,6R)-3,4,5-tris(benzyloxy)-6-(benzyloxymethyl)-tetrahydro-2H-pyran-2-yloxy)octadecan-2-amine

To a mixture of(2S,3R,4S,5S,6R)-2-((2S,3S,4R)-2-azido-3,4-bis(benzyloxy)octadecyloxy)-3,4,5-tris(benzyloxy)-6-(benzyloxymethyl)-tetrahydro-2H-pyran(3.0 g, 2.9 mmol, 1.0 eq) in THF (30 mL) was added 1 M PMe₃ in THF (3.2mL, 3.2 mmol, 1.1 eq) at rt. The mixture was stirred at rt for 5 h, thenH₂O (10 mL) was added, the mixture was stirred at rt for 1 h and thenconcentrated under reduced pressure. The crude residue was purified bycolumn chromatography on silica gel (MeOH/DCM 1:20) to give(2S,3S,4R)-3,4-bis(benzyloxy)-1-((2S,3R,4S,5S,6R)-3,4,5-tris(benzyloxy)-6-(benzyloxymethyl)-tetrahydro-2H-pyran-2-yloxy)octadecan-2-amine(2 g, 68%) as an oil. LC/MS: mass calcd. for C₆₆H₈₅NO₈: 1019.63, found:1020.60 [M+H]⁺.

Synthesis of(2S,3R,4S,5R,6R)-2-(((2S,3S,4R)-2-amino-3,4-dihydroxyoctadecyl)oxy)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol

To a mixture of(2S,3S,4R)-3,4-bis(benzyloxy)-1-(((2S,3R,4S,5S,6R)-3,4,5-tris(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-amine(200 mg, 0.2 mmol, 1.0 eq) in EtOH (10 mL) and DCM (3 mL) was added 20%Pd(OH)₂/C (0.2 g). The mixture was hydrogenated (1 atm) at rt for 16 h,then the catalyst was removed by filtration through a pad of Celite andthe filter cake washed with MeOH. The filtrate was concentrated underreduced pressure to give(2S,3R,4S,5R,6R)-2-(((2S,3S,4R)-2-amino-3,4-dihydroxyoctadecyl)oxy)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol(75 mg, 80%) as an oil.

Synthesis ofN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-11-(3-fluorobicyclo[1.1.1]pentan-1-yl)undecanamideSynthesis of 3-Fluorobicyclo[1.1.1]pentane-1-carbaldehyde

To a mixture of oxalyl chloride (164 mg, 1.29 mmol, 1.5 eq) in DCM (5mL) at −78° C. under an atmosphere of N₂ was added DMSO (202 mg, 2.58mmol, 3.0 eq) dropwise. The mixture was stirred for 15 min at −78° C.then (3-Fluorobicyclo[1.1.1]pentan-1-yl)methanol (100 mg, 0.86 mmol, 1.0eq) in DCM (1 mL) was added dropwise. The mixture was stirred for 50 minat −78° C., then Et₃N (1 mL) was added. Stirring was continued for anadditional 5 min at −78° C., and the mixture was then warmed to rt. H₂O(10 mL) was added and the mixture was extracted with DCM (3×20 mL). Thecombined organic layers were washed with H₂O (20 mL) and brine (20 mL),then dried over Na₂SO₄ and filtered. The filtrate was concentrated underreduced pressure to give 3-fluorobicyclo[1.1.1]pentane-1-carbaldehyde(70 mg, 71%) as an oil. The compound was used without furtherpurification. R_(f)=0.3, PE/EtOAc 1:3.

Synthesis of (10-Carboxydecyl)triphenylphosphonium bromide

To a mixture of 11-bromoundecanoic acid (3.0 g, 11.3 mmol, 1.0 eq) inCH₃CN (100 mL) under an atmosphere of N₂ was added PPh₃ (3.0 g, 11.31mmol, 1.0 eq). The mixture was heated to 90° C., and stirred for 72 h,then concentrated under reduced pressure and the crude productcrystallized from EtOAc give (10-carboxydecyl)triphenylphosphoniumbromide (5.8 g, 97%) as a solid. LC/MS: mass calcd. for C₂₉H₃₆BrO₂P:526, found: 447 [M−Br]⁺.

Synthesis of (E)-11-(3-Fluorobicyclo[1.1.1]pentan-1-yl)undec-10-enoicAcid

To a mixture of (9-carboxynonyl)triphenylphosphonium bromide (315 mg,0.61 mmol, 1.0 eq) in THF (5 mL) at 0° C. under an atmosphere of N₂ wasslowly added NaHMDS, 2 M in THF (0.61 mL, 1.22 mmol, 2 eq). The mixturewas stirred at 0° C. for 1 h, then3-fluorobicyclo[1.1.1]pentane-1-carbaldehyde (70 mg, 0.61 mmol, 1.0 eq)in THF (1 mL) was added at 0° C. The mixture was allowed to warm to rtand stirred for 16 h, then H₂O (10 mL) was added and the pH adjusted to4-5 with 2N HCl. The mixture was extracted with EtOAc (20 mL×3) and thecombined organic layers were concentrated under reduced pressure. Theresidue was purified by column chromatography on silica gel column(PE/EtOAc 2:1) to give(E)-11-(3-fluorobicyclo[1.1.1]pentan-1-yl)undec-10-enoic acid (120 mg,73%) as a solid. LC/MS: mass calcd. for C₁₆H₂₅FO₂: 268, found: 267[M−H]⁻.

Synthesis of 11-(3-Fluorobicyclo[1.1.1]pentan-1-yl)undecanoic Acid

A mixture of (E)-11-(3-fluorobicyclo[1.1.1]pentan-1-yl)undec-10-enoicacid (60 mg, 0.22 mmol, 1.0 eq) and PtO₂ (5 mg, 0.022 mmol, 0.1 eq) inEtOH (50 mL) was hydrogenated (1 atm) at rt for 1 h. The catalyst wasremoved by filtration through a pad of Celite and the filter cake wasrinsed with EtOH. The filtrate was concentrated under reduced pressureto give 11-(3-fluorobicyclo[1.1.1]pentan-1-yl)undecanoic acid (55 mg,91%) as a solid. LC/MS: mass calcd. for C₁₆H₂₇FO₂: 270, found: 269[M−H]⁻.

Synthesis ofN-((2S,3S,4R)-3,4-Dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-11-(3-fluorobicyclo[1.1.1]pentan-1-yl)undecanamide

To a mixture of 11-(3-fluorobicyclo[1.1.1]pentan-1-yl)undecanoic acid(55 mg, 0.20 mmol) and(2S,3R,4S,5R,6R)-2-(((2S,3S,4R)-2-amino-3,4-dihydroxyoctadecyl)oxy)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol(98 mg, 0.20 mmol, 1.0 eq) in THF (5 mL) under an atmosphere of N₂ wasadded HBTU (154 mg, 0.41 mmol, 2.0 eq), Et₃N (41 mg, 0.41 mmol, 2.0 eq),and NMM (41 mg, 0.41 mmol, 2.0 eq) at rt. The mixture was stirred at rtfor 16 h then concentrated under reduced pressure. The residue waspurified by column chromatography on silica (DCM/MeOH 9:1) andpreparative-HPLC to giveN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-11-(3-fluorobicyclo[1.1.1]pentan-1-yl)undecanamide(12.5 mg, 8%) as a solid. LC/MS: mass calcd. for C₄₀H₇₄FNO₉: 731.53,found: 732.45 [M+H]⁺; ¹H NMR (300 MHz, MeOH-d₄) δ 4.21 (d, J=6.0 Hz,1H), 3.81-3.89 (m, 3H), 3.76-3.79 (m, 2H), 3.68-3.74 (m, 3H), 3.62-3.65(m, 1H), 3.53-3.59 (m, 1H), 2.24 (t, J=7.5 Hz, 2H), 1.88 (d, J=2.7 Hz,6H), 1.57-1.66 (m, 6H), 1.29-1.34 (m, 39H), 0.92 (t, J=6.7 Hz, 3H); ¹⁹FNMR (282 MHz, MeOH-d₄) δ−146.5.

Synthesis of (10-Carboxydecyl)triphenylphosphonium Bromide

To a mixture of 11-bromoundecanoic acid (3.0 g, 11.3 mmol, 1.0 eq) inCH₃CN (100 mL) under an atmosphere of N₂ was added PPh₃ (3.0 g, 11.31mmol, 1.0 eq). The mixture was heated to 90° C., and stirred for 72 h,then concentrated under reduced pressure and the crude productcrystallized from EtOAc give (10-carboxydecyl)triphenylphosphoniumbromide (5.8 g, 97%) as a solid. LC/MS: mass calcd. for C₂₉H₃₆BrO₂P:526, found: 447 [M−Br]⁺.

Synthesis of (E)-12(3-Fluorobicyclo[1.1.1]pentan-1-yl)dodec-11-enoicAcid

To a mixture of (10-carboxydecyl)triphenylphosphonium bromide (0.4 g,0.76 mmol, 1.0 eq) in THF (10 mL) at 0° C. under an atmosphere of N₂ wasslowly added NaHMDS, 2 M in THF (0.8 mL, 1.6 mmol, 2.1 eq). The mixturewas stirred at 0° C. for 1 h, then3-fluorobicyclo[1.1.1]pentane-1-carbaldehyde (87 mg, 0.76 mmol, 1.0 eq)in THF (1 mL) was added at 0° C. The mixture was allowed to warm to rtand stirred for 16 h, then H₂O (10 mL) added and the pH adjusted to 4-5with 2N HCl. The mixture was extracted with EtOAc (30 mL×3) and thecombined organic layers were concentrated under reduced pressure. Theresidue was purified by column chromatography on silica gel (PE/EtOAc1:1) to give (E)-12-(3-fluorobicyclo[1.1.1]pentan-1-yl)dodec-11-enoicacid (0.11 g, 51%) as a solid. LC/MS: mass calcd. for C₁₇H₂₇FO₂: 282,found: 281 [M−H]⁻.

Synthesis of 12-(3-Fluorobicyclo[1.1.1]pentan-1-yl)dodecanoic Acid

A mixture of (E)-12-(3-fluorobicyclo[1.1.1]pentan-1-yl)dodec-11-enoicacid (0.11 g, 0.39 mmol, 1.0 eq) and PtO₂ (10 mg, 0.04 mmol, 0.1 eq) inEtOH (70 mL) was hydrogenated (1 atm) at rt for 1 h. The catalyst wasremoved by filtration through a pad of Celite and the filter cake waswashed with EtOH. The filtrate was concentrated under reduced pressureto give 12-(3-fluorobicyclo[1.1.1]pentan-1-yl)dodecanoic acid (0.1 g,90%) as a solid. LC/MS: mass calcd. for C₁₇H₂₉FO₂: 284, found: 283[M−H]⁻.

Synthesis ofN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-21-pyran-2-yl)oxy)octadecan-2-yl)-12-(3-fluorobicyclo[1.1.1]pentan-1-yl)dodecanamide

To a mixture of 12-(3-fluorobicyclo[1.1.1]pentan-1-yl)dodecanoic acid(0.1 g, 0.35 mmol, 1.0 eq) and(2S,3R,4S,5R,6R)-2-(((2S,3S,4R)-2-amino-3,4-bis(benzyloxy)octadecyl)oxy)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol(232 mg, 0.35 mmol, 1.0 eq) in THF (6 mL) under an atmosphere of N₂ wasadded HBTU (267 mg, 0.70 mmol, 2.0 equiv), Et₃N (71 mg, 0.70 mmol, 2.0eq) and NMM (71 mg, 0.70 mmol, 2.0 eq) at rt. The mixture was stirred atrt for 16 h then concentrated under reduced pressure. The residue waspurified by column chromatography on silica gel column (DCM/MeOH 8:1)and preparative-HPLC to giveN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-12-(3-fluorobicyclo[1.1.1]pentan-1-yl)dodecanamide(15.7 mg, 6%) as a sold. LC/MS: mass calcd for C₄₁H₇₆FNO₉: 745.55,found: 746.45 [M+H]⁺; ¹H NMR (300 MHz, MeOH-d₄) δ 4.17 (t, J=5.5 Hz,1H), 3.81-3.89 (m, 3H), 3.74-3.77 (m, 2H), 3.66-3.72 (m, 3H), 3.59-3.63(m, 1H), 3.52-3.57 (m, 1H), 2.22 (t, J=7.4 Hz, 2H), 1.86 (d, J=2.7 Hz,6H), 1.56-1.65 (m, 6H), 1.27-1.35 (m, 41H), 0.95-0.85 (m, 3H); ¹⁹F NMR(282 MHz, MeOH-d₄) δ−146.5.

Synthesis of11-(Bicyclo[2.2.2]octan-1-yl)-N-((2S,3S,4R)-3,4-bis(benzyloxy)-1-(((2S,3R,4S,5S,6R)-3,4,5-tris(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H1-pyran-2-yl)oxy)octadecan-2-yl)undecanamideSynthesis of (E)-11-(Bicyclo[2.2.2]octan-1-yl)undec-10-enoic Acid

To a mixture of 9-carboxynonyl(triphenyl)phosphonium bromide (371 mg,0.72 mmol, 1.0 eq) in THF (5 mL) at 0° C. under an atmosphere of N₂ wasslowly added NaHMDS, 2 M in THF (0.72 mL, 1.44 mmol, 2 eq). The mixturewas stirred at 0° C. for 1 h then bicyclo[2.2.2]octane-1-carbaldehyde(0.1 g, 0.72 mmol, 1.0 eq) in THF (1 mL) was added at 0° C. The mixturewas warmed to rt and stirred for 16 h, then H₂O added and the pHadjusted to ˜4-5 with 2N HCL. The mixture was extracted with EtOAc (30mL×3) and the combined organic layers were concentrated under reducedpressure. The residue was purified by column chromatography on silicagel (PE/EtOAc 2:1) to give(E)-11-(bicyclo[2.2.2]octan-1-yl)undec-10-enoic acid (0.15 g, 71%) as asolid. LC/MS: mass calcd. for C₁₉H₃₂O₂: 292, found: 291 [M−H]⁻.

Synthesis of 11-(Bicyclo[2.2.2]octan-1-yl)undecanoic Acid

A mixture of (E)-11-(bicyclo[2.2.2]octan-1-yl)undec-10-enoic acid (0.15g, 0.51 mmol, 1.0 eq) and 10% Pd/C (20 mg) in MeOH (10 mL) washydrogenated (1 atm) at rt for 1 h. The catalyst was removed byfiltration through a pad of Celite and the filter cake was washed withMeOH. The filtrate was concentrated under reduced pressure to give11-(bicyclo[2.2.2]octan-1-yl)undecanoic acid (130 mg, 86%) as a solid.LC/MS: mass calcd. for C₁₉H₃₄O₂: 294, found: 293 [M−H]⁻.

Synthesis of11-(Bicyclo[2.2.2]octan-1-yl)-N-((2S,3S,4R)-3,4-bis(benzyloxy)-1-(((2S,3R,4S,5S,6R)-3,4,5-tris(benzyloxy-6-((benzyloxy)methyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)undecanamide

To a mixture of 11-(bicyclo[2.2.2]octan-1-yl)undecanoic acid (130 mg,0.44 mmol, 1.0 eq) and(2S,3S,4R)-3,4-bis(benzyloxy)-1-((2S,3R,4S,5S,6R)-3,4,5-tris(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-amine(450 mg, 0.44 mmol, 1.0 eq) in DCM (5 mL) at rt were added EDCI (127 mg,0.66 mmol, 1.5 eq) and DMAP (11 mg, 0.09 mmol, 0.2 eq). The reactionmixture was stirred at rt for 16 h, then diluted with EtOAc (10 mL) andwashed with brine (5 mL). The aqueous layer was extracted with EtOAc (10mL×2) and the combined organic lay ers were dried over Na₂SO₄, filteredand the filtrate concentrated under reduced pressure. The residue waspurified by column chromatography on silica gel (PE/EtOAc 3:1) to give11-(bicyclo[2.2.2]octan-1-yl)-N-((2S,3S,4R)-3,4-bis(benzyloxy)-1-(((2S,3R,4S,5S,6R)-3,4,5-tris(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)undecanamide(0.2 g, 35%) of as a solid. LC/MS: mass calcd. for C₈₅H₁₁₇NO₉: 129%,found: 1297 [M+H]⁺.

Synthesis of11-(bicyclo[2.2.2]octan-1-yl)-N-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)undecanamide

A mixture of11-(bicyclo[2.2.2]octan-1-yl)-N-((2S,3S,4R)-3,4-bis(benzyloxy)-1-(((2S,3R,4S,5S,6R)-3,4,5-tris(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)undecanamide(0.1 g, 0.08 mmol, 1.0 eq) and 20% Pd(OH)₂/C (0.2 g) in EtOH/DCM (10mL/3 mL) was hydrogenated (1 atm) at rt for 16 h. The catalyst wasremoved by filtration through a pad of Celite and the filter cake waswashed with MeOH. The filtrate was concentrated under reduced pressureand the crude residue was purified by column chromatography on silicagel column (DCM/MeOH) and preparative-HPLC to give11-(bicyclo[2.2.2]octan-1-yl)-N-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)undecanamide(8.9 mg, 15%) as a solid. LC/MS: mass calcd. for C₄₃H₈₁NO₉: 755.59,found: 756.50 [M+H]⁺; ¹H NMR (300 MHz, CDCl₃+MeOH-d₄) δ 4.88 (d, J=3.7Hz, 1H), 4.18 (d, J=5.0 Hz, 1H), 3.67-3.89 (m, 8H), 3.53-3.59 (m, 2H),2.20 (t, J=7.7 Hz, 2H), 1.49-1.62 (m, 12H), 1.12-1.30 (m, 43H),0.93-0.99 (m, 2H), 0.87 (t, J=6.4 Hz, 3H).

Synthesis ofN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-11-(2-methylcyclopropyl)undecanamideSynthesis of (E)-11-(2-Methylcyclopropyl)undec-10-enoic Acid

To a mixture of (9-carboxynonyl)triphenylphosphonium bromide (488 mg,0.95 mmol, 1.0 eq) in THF (5 mL) at 0° C. under an atmosphere of N₂ wasslowly added NaHMDS, 2 M in THF (1.0 mL, 2.0 mmol, 2.0 eq). The mixturewas stirred at 0° C. for 1 h, then 2-methylcyclopropane-1-carbaldehyde(80 mg, 0.95 mmol, 1.0 eq) in THF (1 mL) was added at 0° C. The mixturewas warmed to rt and stirred for 16 h, then H₂O (10 mL) added and the pHadjusted to ˜4-5 with 2N HCl. The mixture was extracted with EtOAc (30mL×3) and the combined organic layers were concentrated under reducedpressure. The residue was purified by column chromatography on silicagel (PE/EtOAc 2:1) to give (E)-11-(2-methylcyclopropyl)undec-10-enoicacid (120 mg, 53%) as a solid. LC/MS: mass calcd. for C₁₅H₂₆O₂: 238,found: 237 [M−H]⁻.

Synthesis of 11-(2-Methylcyclopropyl)undecanoic Acid

To a mixture of (E)-11-(2-methylcyclopropyl)undec-10-enoic acid (120 mg,0.50 mmol, 1.0 eq) in THF (5 mL) and H₂O (5 mL) at rt were addedp-MeC₆H₄SO₂NHNH₂ (938 mg, 5.0 mmol, 10 eq) and KOAc (642 mg, 6.6 mmol,13 eq). The mixture was heated to reflux and stirred for 5 h, cooled,and H₂O added. The mixture was extracted with EtOAc (30 mL×3) and thecombined organic layers were concentrated under reduced pressure. Theresidue was purified by column chromatography on silica gel column(PE/EtOAc) to give 11-(2-methylcyclopropyl)undecanoic acid (9) mg, 74%)as an oil. LC/MS: mass calcd. for C₁₅H₂₈O₂: 240, found: 239 [M−H]⁻.

Synthesis ofN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-11-(2-methylcyclopropyl)undecanamide

To a mixture of 11-(2-methylcyclopropyl)undecanoic acid (80 mg, 0.33mmol, 1.0 eq) and(2S,3R,4S,5R,6R)-2-(((2S,3S,4R)-2-amino-3,4-dihydroxyoctadecyl)oxy)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol(160 mg, 0.33 mmol, 1.0 eq) in THF (6 mL) at rt was added HBTU (252 mg,0.66 mmol, 2.0 eq), Et 3N (67 mg, 0.66 mmol, 2.0 eq), and NMM (67 mg,0.66 mmol, 2.0 eq). The mixture was stirred at rt for 16 h, thenconcentrated under reduced pressure and the residue was purified bycolumn chromatography on silica gel (DCM/MeOH 8:1) and preparative-HPLCto giveN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-11-(2-methylcyclopropyl)undecanamide(20.7 mg, 9%) as a solid. LC/MS: mass calcd. for C₃₉H₇₅NO₉: 701.54,found: 702.45 [M+H]⁺; ¹H NMR (300 MHz, MeOH-d₄) δ 4.18 (dd, J=6.5, 4.4Hz, 1H), 3.80-3.89 (m, 3H), 3.74-3.78 (m, 2H), 3.66-3.72 (m, 3H),3.60-3.64 (m, 1H), 3.52-3.57 (m, 1H), 2.22 (t, J=7.5 Hz, 2H), 1.50-1.67(m, 4H), 1.24-1.38 (m, 41H), 1.15-1.23 (m, 2H), 0.99-1.02 (m, 3H),0.95-0.85 (m, 3H), 0.29-0.44 (m, 1H), 0.09-0.18 (m, 1H).

Synthesis of11-(Cuban-1-yl)-N-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)undecanamideSynthesis of (E)-11-Cuban-1-yl)undec-10-enoic Acid

To a mixture of (9-carboxynonyl)triphenylphosphonium bromide (0.4 g, 0.8mmol, 1.0 eq) in THF (6 mL) at 0° C. under an atmosphere of N₂ wasslowly added NaHMDS, 2 M in THF (0.8 mL, 1.6 mmol, 2.0 eq). The mixturewas stirred at 0° C. for 1 h, then cubane-1-carbaldehyde (0.1 g, 0.8mmol, 1.0 eq) in THF (1 mL) was added at 0° C. The mixture was warmed tort and stirred for 16 h, then H₂O (10 mL) added and the pH adjusted to˜4-5 with 2N HCl. The mixture was extracted with EtOAc (20 mL×3) and thecombined organic layers concentrated under reduced pressure. The residuewas purified by column chromatography on silica gel column (PE/EtOAc2:1) to give (E)-11-(cuban-1-yl)undec-10-enoic acid (0.11 g, 49%) as asolid. LC/MS: mass calcd. for C₉₁H₂₆O₂: 286, found 285 [M−H]⁻.

Synthesis of 11-(Cuban-1-yl)undecanoic Acid

A mixture of (E)-11-(cuban-1-yl)undec-10-enoic acid (0.1 g, 0.4 mmol,1.0 eq) and PtO₂ (8 mg, 0.04 mmol, 0.1 eq) in EtOH (10 mL) washydrogenated (1 atm) at rt for 1 h. The catalyst was removed byfiltration through a pad of Celite and the filter cake washed with EtOH.The filtrate was concentrated under reduced pressure to give11-(cuban-1-yl)undecanoic acid (80 mg, 79%) as a solid. LC/MS: masscalcd. for C19H₂₈O₂: 288 found: 287 [M−H]⁻.

Synthesis of11-(Cuban-1-yl)-N-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)undecanamide

To a mixture of 11-(cuban-1-yl)undecanoic acid (80 mg, 0.3 mmol, 1.0 eq)and(2S,3R,4S,5R,6R)-2-(((2S,3S,4R)-2-amino-3,4-dihydroxyoctadecyl)oxy)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol(130 mg, 0.3 mmol, eq) in THF (6 mL) at rt under an atmosphere of N₂ wasadded HBTU (210 mg, 0.55 mmol, 2.0 eq), Et₃N (56 ng, 0.55 mmol, 2.0 eq),and NMM (56 mg, 0.55 mmol, 2.0 eq). The mixture was stirred at rt for 16h then concentrated under reduced pressure. The residue was purified bycolumn chromatography on silica gel (DCM/MeOH) and preparative-HPLC togive11-(cuban-1-yl)-N-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)undecanamideas a solid. LC/MS: mass calcd. for C₄₃H₇₅NO₉: 749.54, found: 750.60[M+H]⁺.

Synthesis of12-(Cuban-1-yl)-N-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)dodecanamideSynthesis of (E)-12-(Cuban-1-yl)dodec-11-enoic Acid

To a mixture of (10-carboxydecyl)triphenylphosphonium bromide (319 mg,0.6 mmol, 1.0 eq) in THF (6 mL) at 0° C. under an atmosphere of N₂ wasslowly added NaHMDS, 2 M in THF (0.6 mL, 1.2 mmol, 2.0 eq). The mixturewas stirred at 0° C. for 1 h, then cubane-1-carbaldehyde (80 mg, 0.6mmol, 1.0 eq) in THF (1 mL) was added. The mixture was warmed to rt andstirred for 16 h, then H₂O (10 mL) added and the pH adjusted to ˜4-5with 2N HCl. The mixture was extracted with EtOAc (20 mL×3) and thecombined organic layers were concentrated under reduced pressure. Theresidue was purified by column chromatography on silica (PE/EtOAc 2:1)to give (E)-12-(cuban-1-yl)dodec-11-enoic acid (110 mg, 60%) as a solid.LC/MS: mass calcd. for C₂₀H₂₈O₂: 300, found: 299 [M−H]⁻.

Synthesis of 12-(Cuban-1-yl)dodecanoic Acid

A mixture of (E)-12-(cuban-1-yl)dodec-11-enoic acid (110 mg, 0.37 mmol,1.0 eq) and PtO₂ (8 mg, 0.04 mmol, 0.1 eq) in EtOH (70 mL) washydrogenated (1 atm) at rt for 1 h. The catalyst was removed byfiltration through a pad of Celite and the filter cake was washed withEtOH. The filtrate was concentrated under reduced pressure to give12-cuban-1-yl)dodecanoic acid (0.1 g, 90%) as a solid. LC/MS: masscalcd. for C₂₀H₃₀O₂: 302, found: 301 [M−H]⁻.

Synthesis of12-(Cuban-1-yl)-N-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)dodecanamide

To a mixture of 12-(cuban-1-yl)dodecanoic acid (80 mg, 0.26 mmol, 1.0eq) and(2S,3R,4S,5R,6R)-2-(((2S,3S,4R)-2-amino-3,4-bis(benzyloxy)octadecyl)oxy)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol(175 mg, 0.26 mmol, 1.0 eq) in THF (6 mL) at rt under an atmosphere ofN₂ was added HBTU (201 mg, 0.53 mmol, 2.0 eq) and TEA (53 mg, 0.53 mmol,2.0 eq) and NMM (53 mg, 0.53 mmol, 2.0 eq). The mixture was stirred atrt for 16 h then concentrated under reduced pressure. The residue waspurified by column chromatography on silica gel (DCM/MeOH) andpreparative-HPLC to give12-(cuban-1-yl)-N-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)dodecanamide(8.6 mg, 4%) as a solid. LC/MS: mass calcd. for C₄₄H₇₇NO₉: 763.56,found: 764.60 [M+H]⁺; ¹H NMR (300 MHz, MeOH-d₄) δ 4.86-4.89 (m, 1H),4.14-4.20 (m, 1H), 4.02-4.08 (m, 1H), 3.80-3.88 (m, 6H), 3.68-3.77 (m,7H), 3.50-3.65 (m, 3H), 2.22 (t, J=7.5 Hz, 2H), 1.48-1.62 (m, 6H),1.27-1.35 (m, 40H), 0.95-0.85 (m, 3H).

Synthesis of(2S,3R,4S,5R,6R)-2-(((2S,3S,4R)-2-(4-(5-(diheptylamino)pentyl)-1H-1,2,3-triazol-1-yl)-3,4-dihydroxyoctadecyl)oxy)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triolSynthesis of N,N-diheptylhept-6-yn-1-amine

To a mixture of hept-6-ynal (0.7 g, 6.4 mmol, 1.0 eq) in DCE (10 mL) atrt under an atmosphere of N₂ was added diheptylamine (1.3 g, 6.4 mmol,1.0 eq). The mixture was stirred at rt for 15 min, then NaBH(OAc)₃ (2.0g, 9.5 mmol, 1.5 eq) and AcOH (0.1 mL) added. The mixture was stirred atrt for 3 h, then H₂O (10 mL) added and the mixture extracted with EtOAc(30 mL×3). The combined organic layers were concentrated under reducedpressure and the residue was purified by column chromatography on silicagel column (MeOH/DCM 1:8) to give N,N-diheptylhept-6-yn-1-amine (0.7 g,36%) as an oil. LC/MS: mass calcd. for C₂₁H₄₁N: 307, found: 308 [M+H]⁺.

Synthesis ofN-(5-(1-((2S,3S,4R)-3,4-bis(benzyloxy)-1-(((2S,3R,4S,5S,6R)-3,4,5-tris(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-1H-1,2,3-triazol-4-yl)pentyl)-N-heptylheptan-1-amine

To a mixture of the N,N-diheptylhept-6-yn-1-amine (38 mg, 0.12 mmol, 1.3eq) and(2S,3R,4S,5S,6R)-2-(((2S,3S,4R)-2-azido-3,4-bis(benzyloxy)octadecyl)oxy)-3,4,5-tris(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran(0.1 g, 0.1 mmol, 1.0 eq) in ^(t)BuOH (3 mL) and H₂O (3 mL) at rt underan atmosphere of N₂ was added CuSO₄ (5 mg, 0.03 mmol, 0.3 eq) and sodiumascorbate (6 mg, 0.03 mmol, 0.3 eq). The mixture was stirred at rt for16 h then diluted with EtOAc (10 mL), washed with brine (5 mL), and theaqueous layer extracted with EtOAc (10 mL×2). The combined organiclayers were dried over Na₂SO₄, filtered and the filtrate concentratedunder reduced pressure. The residue was purified by columnchromatography on silica gel (PE/EtOAc 3:1) to giveN-(5-(1-((2S,3S,4R)-3,4-bis(benzyloxy)-1-(((2S,3R,4S,5S,6R)-3,4,5-tris(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-1H-1,2,3-triazol-4-yl)pentyl)-N-heptylheptan-1-amine(0.1 g, 77%) as an oil. LC/MS: mass calcd. for C₈₇H₁₂₄N₄O₈: 1353, found:1354 [M+H]⁺.

Synthesis of(2S,3R,4S,5R,6R)-2-(((2S,3S,4R)-2-(4-(5-(diheptylamino)pentyl)-1H-1,2,3-triazol-1-yl)-3,4-dihydroxyoctadecyl)oxy)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol

A mixture ofN-(5-(1-((2S,3S,4R)-3,4-bis(benzyloxy)-1-(((2S,3R,4S,5S,6R)-3,4,5-tris(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-1H-1,2,3-triazol-4-yl)pentyl)-N-heptylheptan-1-amine(0.1 g, 0.07 mmol, 1.0 eq) in EtOH (10 mL) and DCM (3 mL) and 20%Pd(OH)₂/C (0.2 g) was hydrogenated (1 atm) at rt for 16 h. The catalystwas removed by filtration through a pad of Celite and the filter cakewashed with EtOH/DCM(3:1). The filtrate was concentrated under reducedpressure and the residue was purified by column chromatography on silicagel (DCM/MeOH 8:1) and preparative-HPLC to give2S,3R,4S,5R,6R)-2-(((2S,3S,4R)-2-(4-(5-(diheptylamino)pentyl)-1H-1,2,3-triazol-1-yl)-3,4-dihydroxyoctadecyl)oxy)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol(16.2 mg, 35%) as a solid. LC/MS: mass calcd. for C₄₅H₈₈N₄O₈: 812.66,found: 813.70 [M+H]⁺; ¹H NMR (300 MHz, DMSO-d₆+D₂O) δ 7.86 (s, 1H),4.90-4.96 (m, 11H), 4.63 (d, J=3.7 Hz, 1H), 4.01-4.06 (m, 1H), 3.87-3.92(m, 1H), 3.47-3.63 (m, 2H), 3.35-3.45 (m, 2H), 3.10-3.18 (m, 1H),2.93-2.99 (m, 6H), 2.55-2.60 (m, 2H), 1.50-1.58 (m, 9H), 1.15-1.26 (m,46H), 0.76-0.83 (m, 9H).

Synthesis of(2S,3R,4S,5R,6R)-2-(((2S,3S,4R)-2-(4-(6-(diheptylamino)hexyl)-1H-1,2,3-triazol-1-yl)-3,4-dihydroxyoctadecyl)oxy)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triolSynthesis of N,N-diheptyloct-7-yn-1-amine

To a mixture of oct-7-ynal (0.7 g, 5.6 mmol, 1.0 eq) in DCE (10 mL) atrt under an atmosphere of N₂ was added diheptylamine (1.2 g, 5.6 mmol,1.0 eq). The mixture was stirred at rt for 15 min, then NaBH(OAc)₃ (1.8g, 8.5 mmol, 1.5 eq) and AcOH (0.1 mL) added. The mixture was stirred atrt for 3 h, then H₂O (10 mL) added and the mixture extracted with EtOAc(30 mL×3). The combined organic layers were concentrated under reducedpressure and the residue was purified by column chromatography on silicagel (MeOH/DCM 1:8) to give N,N-diheptyloct-7-yn-1-amine (0.6 g, 33%) asan oil. LC/MS: mass calcd. for C₂₂H₄₃N: 321, found: 322 [M+H]⁺.

Synthesis ofN-(6-(1-((2S,3S,4R)-3,4-bis(benzyloxy)-1-(((2S,3R,4S,5S,6R)-3,4,5-tris(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-1H-1,2,3-triazol-4-yl)hexyl)-N-heptylheptan-1-amine

To a mixture of the N,N-diheptyloct-7-yn-1-amine (40 mg, 0.12 mmol, 1.0eq) and(2S,3R,4S,5S,6R)-2-(((2S,3S,4R)-2-azido-3,4-bis(benzyloxy)octadecyl)oxy)-3,4,5-tris(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran(130 mg, 0.12 mmol, 1.0 eq) in ^(t)BuOH (3 mL) and H₂O (3 mL) at rt wasadded CuSO₄ (6 mg, 0.04 mmol, 0.3 eq) and sodium ascorbate (7 mg, 0.04mmol, 0.3 eq). The mixture was stirred at rt for 1 day, then dilutedwith EtOAc (10 mL), the mixture washed with brine and the aqueous layerextracted with EtOAc (10 mL×2). The combined organic layers were driedover Na₂SO₄, filtered and concentrated under reduced pressure. Theresidue was purified by column chromatography on silica gel (PE/EtOAc3:1) to giveN-(6-(1-((2S,3S,4R)-3,4-bis(benzyloxy)-1-(((2S,3R,4S,5S,6R)-3,4,5-tris(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-1H-1,2,3-triazol-4-yl)hexyl)-N-heptylheptan-1-amine(0.1 g, 59%) as an oil. LC/MS: mass calcd. for C₈₈H₁₂₆N₄O₈: 1367, found:1368 [M+H]⁺.

Synthesis of(2S,3R,4S,5R,6R)-2-(((2S,3S,4R)-2-(4-(6-(diheptylamino)hexyl)-1H-1,2,3-triazol-1-yl)-3,4-dihydroxyoctadecyl)oxy)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol

A mixture ofN-(6-(1-((2S,3S,4R)-3,4-bis(benzyloxy)-1-(((2S,3R,4S,5S,6R)-3,4,5-tris(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-1H-1,2,3-triazol-4-yl)hexyl)-N-heptylheptan-1-amine(0.1 g, 0.07 mmol, 1.0 eq) in EtOH (10 mL) and DCM (3 mL) and 20%Pd(OH)₂/C (0.2 g) was hydrogenated (1 atm) at rt for 16 h. The catalystwas removed by filtration through a pad of Celite and the filter cakewashed with EtOH/CH₂Cl₂ (3:1). The filtrate was concentrated underreduced pressure and the residue was purified by column chromatographyon silica gel (DCM/MeOH) and preparative-HPLC to give(2S,3R,4S,5R,6R)-2-(((2S,3S,4R)-2-(4-(6-(diheptylamino)hexyl)-1H-1,2,3-triazol-1-yl)-3,4-dihydroxyoctadecyl)oxy)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol(11.1 mg, 18%) as a solid. LC/MS: mass calcd for C₄₆H₉₀N₄O₈: 826.68,found: 827.95 [M+H]⁺; ¹H NMR (300 MHz, DMSO-d₆+D₂O) δ 7.83 (s, 1H),4.88-4.94 (m, 1H), 4.63 (d, J=3.8 Hz, 1H), 3.97-4.12 (m, 1H), 3.86-3.93(m, 1H), 3.34-3.53 (m, 5H), 3.08-3.14 (m, 1H), 2.93-3.00 (m, 6H),2.54-2.60 (m, 2H), 1.43-1.59 (m, 10H), 1.18-1.27 (m, 46H), 0.80-0.85 (m,9H).

Synthesis ofN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-21-pyran-2-yl)oxy)octadecan-2-yl)dodecane-1-sulfinamide—Mixtureof Two Diastereomers

Synthesis of S-dodecyl Ethanethioate

To a mixture of 1-bromododecane (0.5 g, 2.0 mmol) in THF (10 mL) at rtwas added potassium ethanethioate (274 mg, 2.4 mmol). The mixture washeated to 80° C., and stirred for 3 h, then concentrated under reducedpressure and the residue was purified by column chromatography on silicagel (PE/EtOAc) to give S-dodecyl ethanethioate (382 mg, 78%) as an oil.

Synthesis of dodecane-1-sulfinic Chloride

To a mixture of S-dodecyl ethanethioate (0.3 g, 1.2 mmol) in DCM (5 mL)at −20° C. was slowly added Ac₂O (126 mg, 1.2 mmol) and SO₂Cl₂ (332 mg,2.5 mmol). The mixture was warmed to −5° C., and stirred for 2 h, thenconcentrated under reduced pressure to give dodecane-1-sulfinic chloride(295 mg, 95%) as an oil.

Synthesis ofN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)dodecane-1-sulfinamide—Mixtureof Two Diastereomers

To a mixture of(2S,3R,4S,5R,6R)-2-(((2S,3S,4R)-2-amino-3,4-dihydroxyoctadecyl)oxy)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol(60 mg, 0.13 mmol) in DMA (3 mL) at rt under an atmosphere of N₂ wasadded Et₃N (25 mg, 0.25 mmol) and dodecane-1-sulfinic chloride (47 mg,0.19 mmol). The mixture was stirred at rt for 3 h. then H₂O added andthe mixture was extracted with EtOAc (3×30 mL). The combined organiclayers were dried over anhydrous Na₂SO₄ and filtered. The filtrate wasconcentrated under reduced pressure and the residue was purified twiceby column chromatography on silica gel (DCM/MeOH 9:1) to giveN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)dodecane-1-sulfinamide(3.4 mg, 3.9%) as a solid. LC/MS: mass calcd. for C₃₆H₇₃NO₉S: 695.50,found: 696.55 [M+H]⁺.

Synthesis ofN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-10-(3-fluorobicyclo[1.1.1]pentan-1-yl)decane-1-sulfinamide—Mixtureof Two Diastereomers

Synthesis of 9-(bromotriphenyl-lambda5-phosphanyl)nonanoic Acid

A mixture of 9-bromononanoic acid (5.0 g, 21.1 mmol) and Ph₃P (5.53 g,21.1 mmol) in MeCN (50 mL) was heated to reflux and stirred for 3 days.The mixture was concentrated under reduced pressure and the residue wastriturated with mixture Et₂O (200 mL) and filtered to give9-(bromotriphenyl-lambda5-phosphanyl)nonanoic acid (10.0 g, 95%) as asolid. ¹H NMR (300 MHz, DMSO-d₆) δ 11.98 (s, 1H), 7.42-8.28 (m, 15H),2.17 (t, J=7.3 Hz, 2H), 1.76 (td, J=6.6, 5.8, 2.5 Hz, 2H), 1.39-1.56 (m,6H), 1.14-1.29 (m, 6H).

Synthesis of (9E)-10-[3-fluorobicyclo[1.1.1]pentan-1-yl]dec-9-enoic Acid

To a mixture of 9-(bromotriphenyl-lambda5-phosphanyl)nonanoic acid (2.95g, 5.9 mmol) in THF (50 mL) at −10° C. under an atmosphere of N₂ wasadded 2 M NaHMDS (5.9 mL, 11.8 mmol). The mixture was warmed to rt andstirred for 1 h at room temperature, then a mixture of3-fluorobicyclo[1.1.1]pentane-1-carbaldehyde (450 mg, 3.9 mmol) in THF(5 mL) was added. The mixture was stirred at rt overnight, then 2 M HCl(20 mL) added and the mixture extracted with EtOAc (3×20 mL). Thecombined organic layers were washed with brine (50 mL), dried andfiltered. The filtrate was concentrated under reduced pressure and theresidue was purified by column chromatography on silica gel (PE/EtOAc1:1) to give (9E)-10-[3-fluorobicyclo[1.1.1]pentan-1-yl]dec-9-enoic acid(500 mg, 50%) as an oil LC/MS: mass calcd. for C₁₅H₂₃FO₂: 254.2, found:253.9 [M−H]⁻.

Synthesis of 10-[3-fluorobicyclo[1.1.1]pentan-1-yl]decanoic Acid

A mixture of (9E)-10-[3-fluorobicyclo[1.1.1]pentan-1-yl]dec-9-enoic acid(500 mg, 1.96 mmol), and PtO₂ (50 mg, 0.22 mmol) in EtOH (30) mL) wasstirred under an atmosphere of H₂ (balloon) for 2 h. The mixture wasfiltered, the filter cake was washed with EtOH (100 mL) and the filtratewas concentrated under reduced pressure to give10-[3-fluorobicyclo[1.1.1]pentan-1-yl]decanoic acid (500 mg, 99%) as asolid. LC/MS: mass calcd. for C₁₅H₂₅FO₂: 256.2, found: 255.0 [M−H]⁻.

Synthesis of 10-{3-fluorobicyclo[1.1.1]pentan-1-yl}decan-1-ol

To a mixture of 10-{3-fluorobicyclo[1.1.1]pentan-1-yl}decanoic acid (500mg, 1.95 mmol) in THF (20 mL) under an atmosphere of N₂ was added 1 MBH₃ in THF (5.9 mL, 5.9 mmol) dropwise. The mixture was stirred at rtfor 2 h, then quenched with MeOH (20 mL), concentrated under reducedpressure and the residue was purified by column chromatography on silica(PE/EtOAc 3:1) to give 10-{3-fluorobicyclo[1.1.1]pentan-1-yl}decan-1-ol(430 mg, 91%) as an oil. ¹H NMR (300 MHz, DMSO-d₆) δ 4.32 (t, J=5.2 Hz,1H), 3.37 (td, J=6.5, 5.1 Hz, 2H), 1.88 (d, J=2.7 Hz, 6H), 1.60 (d,J=7.8 Hz, 2H), 1.40 (t, J=6.5 Hz, 2H), 1.25 (s, 12H).

Synthesis of 1-(10-bromodecyl)-3-fluorobicyclo[1.1.1]pentane

To a mixture of 10-{3-fluorobicyclo[1.1.1]pentan-1-yl}decan-1-ol (430mg, 1.77 mmol) in DCM (30 mL) under an atmosphere of N₂ was added Ph₃P(930 mg, 3.54 mmol) and CBr₄ (1.18 g, 3.54 mmol). The mixture wasstirred at rt for 4 h, then concentrated under reduced pressure and theresidue was purified by column chromatography on silica gel (PE/EtOAc10:1) to give 1-(10-bromodecyl)-3-fluorobicyclo[1.1.1]pentane (480 mg,89%) as an oil. ¹H NMR (400 MHz, DMSO-d₆) δ 3.53 (t, J=6.7 Hz, 2H), 1.88(d, J=2.8 Hz, 61), 1.79 (d, J=6.8 Hz, 2H), 1.59 (t, J=7.0 Hz, 2H),1.33-1.41 (m, 2H), 1.26 (d, J=1.7 Hz, 12H).

Synthesis of1-[(10-{3-fluorobicyclo[1.1.1]pentan-1-yl}decyl)sulfanyl]ethanone

A mixture of 1-(10-bromodecyl)-3-fluorobicyclo[1.1.1]pentane (480 mg,1.57 mmol) and 1-(potassiosulfanyl)ethanone (359 mg, 3.14 mmol) in THF(20 mL) was heated to 60° C. and stirred for 4 h, then concentratedunder reduced pressure and the residue was purified by columnchromatography on silicagel (PE/EtOAc 10:1) to give1-[(10-{3-fluorobicyclo[1.1.1]pentan-1-yl}decyl)sulfanyl]ethanone (450mg, 95%) as an oil. ¹H NMR (300 MHz, CDCl₃) δ 2.88 (t, J=7.3 Hz, 2H),2.35 (s, 3H), 1.90 (d, J=2.7 Hz, 6H), 1.50-1.67 (m, 4H), 1.28 (d, J=2.7Hz, 14H).

Synthesis of 10-{3-fluorobicyclo[1.1.1]pentan-1-yl}decane-1-sulfinylChloride

To a mixture of1-[(10-{3-fluorobicyclo[1.1.1]pentan-1-yl}decyl)sulfanyl]ethanone (50mg, 0.17 mmol) in DCM (0.5 mL) under an atmosphere of N₂ at −20° C. wasadded Ac₂O (17 mg, 0.17 mmol) and SO₂Cl₂ (45 mg, 0.33 mmol). The mixturewas stirred at −20° C. for 10 min, then concentrated under reducedpressure to give 10-{3-fluorobicyclo[1.1.1]pentan-1-yl}decane-1-sulfinylchloride (50 mg, 97%) as an oil.

Synthesis ofN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-10-(3-fluorobicyclo[1.1.1]pentan-1-yl)decane-1-sulfinamide—Mixtureof Two Diastereomers

To a mixture of(2S,3R,4S,5R,6R)-2-{[(2S,3S,4R)-2-amino-3,4-dihydroxyoctadecyl]oxy}-6-(hydroxymethyl)oxane-3,4,5-triol(50 mg, 0.10 mmol) in DMA (3 mL) and DCM (1 mL) was added Et₃N (106 mg,1.0 mmol) and 10-(3-fluorobicyclo[1.1.1]pentan-1-yl)decane-1-sulfinylchloride (50 mg, 0.17 mmol). The mixture was stirred at rt for 1 h, thenconcentrated under reduced pressure and the residue was purified bypreparative-HPLC to giveN-((2S,3S,4R)-3,4-dihydroxy-1-((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-10-(3-fluorobicyclo[1.1.1]pentan-1-yl)decane-1-sulfinamide—mixtureof two diastereomers (5.1 mg, 7%) as a solid. LC/MS: mass calcd. forC₃₉H₇₄FNO₉S: 751.51, found: 752.07 [M+H]⁺; ¹H NMR (400 MHz, CD₃OD) δ4.10 (dd, J=9.5, 2.2 Hz, 1H), 3.85-3.98 (m, 2H), 3.65-3.85 (m, 4H),3.55-3.65 (m, 2H), 3.45-3.55 (m, 1H), 2.89 (qt, J=13.0, 7.4 Hz, 2H),1.88 (d, J=2.6 Hz, 6H), 1.54-1.73 (m, 6H), 1.23-1.32 (m, 40H), 0.90-0.95(m, 3H).

Synthesis ofN-[(2S,3S,4R)-3,4-dihydroxy-1-[(2S,3S)-2,3,4-trihydroxybutoxy]octadecan-2-yl]-11-[3-fluorobicyclo[1.1.1]pentan-1-yl]undecanamide

To a mixture of(2S,3S)-4-[[(2S,3S,4R)-2-amino-3,4-dihydroxyoctadecyl]oxy]butane-1,2,3-triol[Diazet al Tetrahedron: Asymmetry 2009, 20, 747-753 and Jervis et alBioconjugate Chemistry 2013, 24, 586-594] (50 mg, 0.12 mmol) and11-[3-fluorobicyclo[1.1.1]pentan-1-yl]undecanoic acid (35.3 mg, 0.13mmol) in DMF (3 mL) at rt was added HBTU (90 mg, 0.24 mmol). Et₃N (24mg, 0.24 mmol) and NMM (24 mg, 0.24 mmol). The mixture was stirred at rtovernight, then purified by preparative-HPLC to affordN-[(2S,3S,4R)-3,4-dihydroxy-1-[(2S,3S)-2,3,4-trihydroxybutoxy]octadecan-2-yl]-11-[3-fluorobicyclo[1.1.1]pentan-1-yl]undecanamide(7.8 mg, 10%) as a solid LC/MS: mass calcd. for C₃₈H₇₂FNO₇: 673.53,found: 674.50 [M+H]⁺; ¹H NMR (300 MHz, CD₃OD) δ 4.20 (s, 1H), 3.74-3.84(m, 1H), 3.52-3.72 (m, 8H), 3.20-3.30 (m, 1H), 2.23 (t, J=7.5 Hz, 2H),1.88 (s, 5H), 1.28-1.63 (m, 45H), 0.92 (t, J=6.4 Hz, 3H).

Synthesis of((2R,3R,4S,5R,6S)-6-(((2S,3S,4R)-2-(11-(3-fluorobicyclo[1.1.1]pentan-1-yl)undecenamido)-3,4-dihydroxyoctadecyl)oxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-yl)methyl((4R)-4-((3R,7R,10S,13R)-3,7-dihydroxy-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-17-yl)pentanoyl)-L-valinate

Synthesis of((2R,3S,4S,5R,6S)-6-(((2S,3S,4R)-2-azido-3,4-bis(benzyloxy)octadecyl)oxy)-3,4,5-tris(benzyloxy)tetrahydro-2H-pyran-2-yl)methyl(((9H-fluoren-9-yl)methoxy)carbonyl)-L-valinate

To a mixture of((2R,3S,4S,5R,6S)-6-(((2S,3S,4R)-2-azido-3,4-bis(benzyloxy)octadecyl)oxy)-3,4,5-tris(benzyloxy)tetrahydro-2H-pyran-2-yl)methanol[Org. Biomol. Chem. 2011, 9, 8413] (0.7 g, 0.7 mmol) and(((9H-fluoren-9-yl)methoxy)carbonyl)-L-valine (0.5 g, 1.5 mmol) in THF(20 mL) at rt was added EDCI (210 mg, 1.10 mmol) and DMAP (179 mg, 1.46mmol). The mixture was stirred for 16 h at rt, then concentrated undervacuum and the residue purified by silica gel column chromatography(PE/EtOAc 3:1) to give((2R,3S,4S,5R,6S)-6-(((2,S,3S,4R)-2-azido-3,4-bis(benzyloxy)octadecyl)oxy)-3,4,5-tris(benzyloxy)tetrahydro-2H-pyran-2-yl)methyl(((9H-fluoren-9-yl)methoxy)carbonyl)-L-valinate (0.8 g, 86%) as an oil.LC/MS: mass calcd. for C₇₉H₉₆N₄O₁₁: 1277, found: 1278 [M+H]⁺.

Synthesis of((2R,3S,4S,5R,6S)-6-(((2S,3S,4R)-2-azido-3,4-bis(benzyloxy)octadecyl)oxy)-3,4,5-tris(benzyloxy)tetrahydro-2H-pyran-2-yl)methylL-valinate

To a mixture of((2R,3S,4S,5R,6S)-6-(((2S,3S,4R)-2-azido-3,4-bis(benzyloxy)octadecyl)oxy)-3,4,5-tris(benzyloxy)tetrahydro-2H-pyran-2-yl)methyl(((9H-fluoren-9-yl)methoxy)carbonyl)-L-valinate (0.8 g, 0.6 mmol) in DMF(15 mL) at was added piperidine (0.3 g, 3.5 mmol). The mixture wasstirred at rt for 0.5 h, then was quenched with H—O and extracted withEtOAc. The combined organic layers were washed with brine, dried overNa₂SO₄ and filtered. The filtrate was concentrated under reducedpressure and the residue was purified by silica gel column (PE/EtOAc2:1) to give((2R,3S,4S,5R,6S)-6-(((2S,3S,4R)-2-azido-3,4-bis(benzyloxy)octadecyl)oxy)-3,4,5-tris(benzyloxy)tetrahydro-2H-pyran-2-yl)methylL-valinate (0.6 g, 91%) as an oil. LC/MS: mass calcd. for C₆₄H₈₆N₄O₉:1055, found: 1056 [M+H]⁺.

Synthesis of((2R,3S,4S,5R,6S)-6-(((2S,3S,4R)-2-azido-3,4-bis(benzyloxy)octadecyl)oxy)-3,4,5-tris(benzyloxy)tetrahydro-2H-pyran-2-yl)methyl((4R)-4-((3R,7R,10S,13R)-3,7-dihydroxy-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-17-yl)pentanoyl)-L-valinate

To a mixture of((2R,3S,4S,5R,6S)-6-(((2S,3S,4R)-2-azido-3,4-bis(benzyloxy)octadecyl)oxy)-3,4,5-tris(benzyloxy)tetrahydro-2H-pyran-2-yl)methylL-valinate (0.6 g, 0.6 mmol) and chenodeoxycholic acid (268 mg, 0.68mmol) in DMF (10 mL) at rt was added HATU (324 mg, 0.85 mmol) and DIPEA(147 mg, 1.14 mmol). The mixture was stirred at rt for 6 h then H₂Oadded and the mixture extracted with EtOAc. The combined organic layerswere washed with brine, dried over Na₂SO₄ and filtered. The filtrate wasconcentrated under reduced pressure and the residue was purified bysilica gel column chromatography (PE/EtOAc 3:1) to give((2R,3S,4S,5R,6S)-6-(((2S,3S,4R)-2-azido-3,4-bis(benzyloxy)octadecyl)oxy)-3,4,5-tris(benzyloxy)tetrahydro-2H-pyran-2-yl)methyl((4R)-4-((3R,7R,10S,13R)-3,7-dihydroxy-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-17-yl)pentanoyl)-L-valinate(0.6 g, 74%) as an oil. LC/MS: mass calcd. for C₈₈H₁₂₄N₄O₁₂: 1429,found: 1430 [M+H]⁺.

Synthesis of((2R,3R,4S,5R,6S)-6-(((2S,3S,4R)-2-amino-3,4-dihydroxyoctadecyl)oxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-yl)methyl((4R)-4-((3R,7R,10S,13R)-3,7-dihydroxy-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-17-yl)pentanoyl)-L-valinate

A mixture of((2R,3S,4S,5R,6S)-6-(((2S,3S,4R)-2-azido-3,4-bis(benzyloxy)octadecyl)oxy)-3,4,5-tris(benzyloxy)tetrahydro-2H-pyran-2-yl)methyl((4R)-4-((3R,7R,10S,13R)-3,7-dihydroxy-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-17-yl)pentanoyl)-L-valinate(0.6 g, 0.4 mmol) and 20% Pd(OH)₂/C (0.6 g) in MeOH (15 mL) and DCM (15mL) was hydrogenated (1 atm) at rt for 16 h. The mixture was filteredthrough a pad of Celite and the filter cake was washed with MeOH. Thefiltrate was concentrated under reduced pressure to give((2R,3R,4S,5R,6S)-6-(((2S,3S,4R)-2-amino-3,4-dihydroxyoctadecyl)oxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-yl)methyl((4R)-4-((3R,7R,10S,13R)-3,7-dihydroxy-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-17-yl)pentanoyl)-L-valinate(0.33 g, 83%) as a solid. LC/MS: mass calcd. for C₅₃H₉₆N₂O₁₂: 953,found: 954 [M+H]⁺.

Synthesis of((2R,3R,4S,5R,6S)-6-(((2S,3S,4R)-2-(11-(3-fluorobicyclo[1.1.1]pentan-1-yl)undecenamido)-3,4-dihydroxyoctadecyl)oxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-yl)methyl((4R)-4-((3R,7R,10S,13R)-3,7-dihydroxy-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-17-yl)pentanoyl)-L-valinate

To a mixture of((2R,3R,4S,5R,6S)-6-(((2S,3S,4R)-2-amino-3,4-dihydroxyoctadecyl)oxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-yl)methyl((4R)-4-((3R,7R,10S,13R)-3,7-dihydroxy-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-17-yl)pentanoyl)-L-valinate(100 mg, 0.11 mmol) and 11-(3-fluorobicyclo[1.1.1]pentan-1-yl)undecanoicacid (31 mg, 0.12 mmol) in THF (5 mL) and DMF (5 mL) at rt under anatmosphere of N₂ was added HBTU (119 mg, 0.32 mmol), Et₃N (0.1 mL) andNMM (0.1 mL). The mixture was stirred at rt for 16 h, then quenched withH₂O and extracted with EtOAc. The combined organic layers were washedwith brine, dried over Na₂SO₄ and filtered. The filtrate was purified bysilica gel column chromatography (DCM/MeOH) to give a crude product (50mg), which was purified further by trituration with CH₃CN to give((2R,3R,4S,5R,6S)-6-(((2S,3S,4R)-2-(l1-(3-fluorobicyclo[1.1.1]pentan-1-yl)undecenamido)-3,4-dihydroxyoctadecyl)oxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-yl)methyl((4R)-4-((3R,7R,10S,13R)-3,7-dihydroxy-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-17-yl)pentanoyl)-L-valinate(26 mg, 21%) as a solid. LC/MS: mass calcd. for C₆₉H₁₂₁FN₂O₁₃: 1205,found: 1206 [M+H]⁺.

Synthesis ofN-[(2S,3S,4R)-3,4-dihydroxy-1-{[(2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy}octadecan-2-yl]-11-(oxetan-3-yl)undecanamide

Step 1: Synthesis of 11-(oxetan-3-yl)undec-10-enoic Acid

To a mixture of (9-carboxynonyl)triphenylphosphonium bromide (565 mg,1.1 mmol) in THF (20 mL) at 0° C. under an atmosphere of N₂ was addedNaHMDS, 2.0 M (1.1 mL, 2.2 mmol). The mixture was warmed to roomtemperature and stirred for 1 h, then oxetane-3-carbaldehyde (86 mg, 1.0mmol) in THF (1 mL) was added. The mixture was stirred overnight at rt,then acidified pH˜1 with 1N HCl and extracted with EtOAc (3×30 mL). Thecombined organic layers were washed with brine (30 mL), dried overanhydrous Na₂SO₄, filtered, and concentrated under reduced pressure. Theresidue was purified by column chromatography on silica gel (PE/EtOAc1:1 to give 11-(oxetan-3-yl)undec-10-enoic acid (100 mg, 42%) as an oil.LC/MS: mass calcd. for C₁₄H₂₄O₃: 240, found: 239 [M−H]⁻.

Step 2: Synthesis of 11-(oxetan-3-yl)undecanoic

A mixture of 11-(oxetan-3-yl)undec-10-enoic acid (100 mg, 0.4 mmol),PtO₂ (20 mg. 0.1 mmol) and EtOH (30 mL) was stirred under an atmosphereof H₂ (balloon) for 1 h. The mixture was filtered through a pad ofCelite and the filtrate was concentrated under reduced pressure to give11-(oxetan-3-yl)undecanoic acid (100 mg, 99%) as a solid. LC/MS: masscalcd. for C₁₄H₂₆O₃: 242, found: 241 [M−H]⁻.

Step 3: Synthesis ofN-[(2S,3S,4R)-3,4-dihydroxy-1-{[(2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy}octadecan-2-yl]-11-(oxetan-3-yl)undecanamide

To a mixture of(2S,3R,4S,5R,6R)-2-(((2S,3S,4R)-2-amino-3,4-dihydroxyoctadecyl)oxy)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol(50 mg, 0.1 mmol) in DMF (2 mL) under an atmosphere of N₂ was added11-(oxetan-3-yl)undecanoic acid (25 mg, 0.1 mmol), Et₃N (0.1 mL), NMM(0.1 mL) and HBTU (80 mg, 0.2 mmol). The mixture was stirred at rt for16 h, then diluted with H₂O (10 mL) and extract with EtOAc (30 mL×3).The combined organic layers were washed with brine (30 mL×2), dried overNa₂SO₄ and filtered. The filtrate was concentrated under reducedpressure and the residue was purified by column chromatography on silicagel (DCM/MeOH 5:1) and preparative-HPLC to giveN-[(2S,3S,4R)-3,4-dihydroxy-1-{[(2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy}octadecan-2-yl]-11-(oxetan-3-yl)undecanamide(13.8 mg, 19%) as a solid. LC/MS: mass calcd. for C₃₈H₇₃NO₁₀: 703.99,found: 704.50 [M+H]⁺. ¹H NMR (300 MHz, CD₃OD) δ 4.81 (dd, J=7.9, 5.8 Hz,2H), 4.39 (t, J=6.1 Hz, 2H), 4.21 (d, J=5.7 Hz, 1H), 3.56-3.91 (m, 10H),2.99-3.01 (m, 1H), 2.35-2.18 (m, 2H), 1.63-1.70 (m, 6H), 1.33-1.46 (m,39H), 0.87-0.97 (m, 3H).

Synthesis ofN-[(2S,3S,4R)-3,4-dihydroxy-1-{[(2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy}octadecan-2-yl]-11-(3-methyloxetan-3-yl)undecanamide

Step 1: Synthesis of 11-(3-methyloxetan-3-yl)undec-10-enoic Acid

To a mixture of (9-carboxynonyl)triphenylphosphonium bromide (564 mg,1.1 mmol) in THF (20 mL) at 0° C. under an atmosphere of N₂ was added 2M NaHMDS (1.1 mL, 2.2 mmol). The mixture was warmed to rt and stirredfor 1 h, then 3-methyloxetane-3-carbaldehyde (100 mg, 1.0 mmol) in THF(2 mL) was added. The mixture was stirred at rt overnight, thenacidified to pH˜1 with 1N HCl, and extracted with EtOAc (3×20 mL). Thecombined organic layers were washed with brine (30 mL), dried overanhydrous Na₂SO₄, filtered, and concentrated under reduced pressure. Theresidue was purified by column chromatography on silica gel (PE/EA 1:1)to give 11-(3-methyloxetan-3-yl)undec-10-enoic acid (110 mg, 43%) as anoil LC/MS: mass calcd. for C₁₅H₂₆O₃: 254, found: 253 [M−H]⁻.

Step 2: Synthesis of 11-(3-methyloxetan-3-yl)undecanoic Acid

A mixture of 11-(3-methyloxetan-3-yl)undec-10-enoic acid (110 mg, 0.4mmol) and PtO₂ (20 mg, 0.1 mmol) in EtOH (30 mL) was stirred under anatmosphere of H₂ (balloon) for 1 h. The mixture was filtered through apad of Celite and the filtrate was concentrated under reduced pressureto give 11-(3-methyloxetan-3-yl)undecanoic acid (110 mg, 99%) as asolid. LC/MS: mass calcd. for C₁₅H₂₈O₃: 256, found: 255 [M−H]⁻.

Step 3: Synthesis ofN-[(2S,3S,4R)-3,4-dihydroxy-1-{[(2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy}octadecan-2-yl]-11-(3-methyloxetan-3-yl)undecanamide

To a mixture of(2S,3R,4S,5R,6R)-2-(((2S,3S,4R)-2-amino-3,4-dihydroxyoctadecyl)oxy)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol(50 mg, 0.1 mmol) in DMF (2 mL) under an atmosphere of N₂ was added11-(3-methyloxetan-3-yl)undecanoic acid (27 mg, 0.1 mmol), Et₃N (0.1mL), NMM (0.1 mL) and HBTU (80 mg, 0.2 mmol). The mixture was stirred atrt for 16 h, then diluted with H₂O (10 mL) and extract with EtOAc (30mL×3). The combined organic layers were washed with brine (30 mL×2),dried over Na₂SO₄, filtered and the filtrate was concentrated underreduced pressure. The residue was purified by column chromatography onsilica gel (DCM/MeOH 5.1) and preparative-HPLC to giveN-[(2S,3S,4R)-3,4-dihydroxy-1-{[(2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy}octadecan-2-yl]-11-(3-methyloxetan-3-yl)undecanamide(10.2 mg, 13%) as a solid. LC/MS: mass calcd. for C₃₉H₇₅NO₁₀: 718.01,found: 718.50 [M+H]⁺; ¹H NMR (300 MHz, CD₃OD) δ 4.44 (d, J=0.1=5.6 Hz,2H), 4.35 (d, J=5.5 Hz, 2H), 4.15-4.25 (m, 1H), 3.53-3.83 (m, 10H),3.31-3.32 (m, 1H), 2.24 (t, J=7.5 Hz, 2H), 1.52-1.65 (m, 6H), 1.28-1.36(m, 41H), 0.83-0.97 (m, 3H).

Synthesis of11-((1r,3s)-adamantan-1-yl)-N-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)undecanamide

Step 1: Synthesis of (11-((1s,3s)-adamantan-1-yl)undec-10-enoic Acid

Prepared in a manner similar to 11-(3-methyloxetan-3-yl)undec-10-enoicacid and purified by column chromatography on silica gel (PE/EtOAc 1:1)to give (11-((1s,3s)-adamantan-1-yl)undec-10-enoic acid (250 mg, 799%)as a solid. LC/MS: mass calcd. for C₂₁H₃₄O₂: 318, found: 317 [M−H]⁻.

Step 2: Synthesis of 11-((1r,3s)-adamantan-1-yl)undecanoic Acid

A mixture of (11-((1s,3s)-adamantan-1-yl)undec-10-enoic acid (250 mg,0.7 mmol) and PtO₂ (40 mg, 0.2 mmol) in EtOH (30 mL) was stirred underan atmosphere of H₂ (balloon) for 1 h. The mixture was filtered througha pad of Celite and the filtrate was concentrated under reduced pressureto give 11-((1r,3s)-adamantan-1-yl)undecanoic acid (250 mg, 99.4%) as asolid. LC/MS: mass calcd. for C₂₁H₃₆O₂: 320, found: 319 [M−H]⁻.

Step 3: Synthesis of11-((1r,3s)-adamantan-1-yl)-N-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)undecanamide

Prepared in a manner similar toN-[(2S,3S,4R)-3,4-dihydroxy-1-{[(2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy}octadecan-2-yl]-11-(3-methyloxetan-3-yl)undecanamide,purified by column chromatography on silica gel (DCM/MeOH 5:1) andpreparative-HPLC to give11-((1r,3s)-adamantan-1-yl)-N-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)undecanamide(5.4 mg, 6.4%) as a solid. LC/MS: mass calcd. for C₄₅H₈₃NO₉: 782.14,found: 782.65 [M+H]⁺; ¹H NMR (300 MHz, CD₃OD) δ 4.19 (dd, J=6.7, 4.4 Hz,1H), 3.54-3.94 (m, 10H), 3.31-3.32 (m, 1H), 2.33 (t, J=7.4 Hz, 1H), 2.24(t, J=7.4 Hz, 1H), 1.99-1.91 (m, 4H), 1.65-1.86 (m, 11H), 1.49-1.53 (m,7H), 1.26-1.36 (m, 34H), 1.05-1.09 (m, 3H), 0.87-0.91 (m, 3H).

Synthesis ofN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-11-(tetrahydrofuran-3-yl)undecanamide

Step 1: Synthesis of 11-(tetrahydrofuran-3-yl)undec-10-enoic Acid

Prepared in a manner similar to 11-(3-methyloxetan-3-yl)undec-10-enoicacid and purified by column chromatography on silica gel (PE/EtOAc 1:1)to give 11-(tetrahydrofuran-3-yl)undec-10-enoic acid (250 mg, 79%) assolid. LC/MS: mass calcd. for C₁₅H₂₆O₃: 254, found: 253 [M−H]⁻.

Step 2: Synthesis of 11-(tetrahydrofuran-3-yl)undecanoic Acid

A mixture of 11-(tetrahydrofuran-3-yl)undec-10-enoic acid (200 mg, 0.7mmol) and PtO₂ (40 mg, 0.2 mmol) in EtOH (30 mL) was stirred under anatmosphere of H₂ (balloon) for 1 h, then filtered through a pad ofCelite and the filtrate was concentrated under reduced pressure to give11-(tetrahydrofuran-3-yl)undecanoic acid (200 mg, 99%) as a solid.LC/MS: mass calcd. for C₁₅H₂₈O₃: 256, found: 255 [M−H]⁻.

Step 3: Synthesis ofN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-11-(tetrahydrofuran-3-yl)undecanamide

Prepared in a manner similar toN-[(2S,3S,4R)-3,4-dihydroxy-1-{[(2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy}octadecan-2-yl]-11-(3-methyloxetan-3-yl)undecanamide,purified by column chromatography on silica gel (DCM/MeOH 5:1) andpreparative-HPLC to giveN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-11-(tetrahydrofuran-3-yl)undecanamide(9 mg, 8%) as a solid. LC/MS: mass calcd. for C₃₉H₇₅NO₁₀: 717.54, found:718.60 [M+H]⁺; ¹H NMR (300 MHz, MeOHl-d₄) δ 4.21 (d, J=5.7 Hz, 1H),3.56-3.92 (m, 12H), 2.15-2.20 (m, 2H), 1.63-1.70 (m, 5H), 1.33-1.46 (s,44H), 0.87-0.97 (m, 3H).

Synthesis ofN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-11-(tetrahydro-2H-pyran-4-yl)undecanamide

Step 1: Synthesis of 11-(tetrahydro-2H-pyran-4-yl)undec-10-enoic Acid

Prepared in a manner similar to 11-(3-methyloxetan-3-yl)undec-10-enoicacid and purified by column chromatography on silica gel (PE/EtOAc 1:1)to give 11-(tetrahydro-2H-pyran-4-yl)undec-10-enoic acid (200 mg, 85%)as a solid. LC/MS: mass calcd. for C₁₆H₂₈O₃: 268, found: 267 [M−H]⁻.as asolid.

Step 2: Synthesis of 11-(tetrahydro-2H-pyran-4-yl)undecanoic Acid

A mixture of 11-(tetrahydro-2H-pyran-4-yl)undec-10-enoic acid (200 mg,0.7 mmol) and PtO₂ (40 mg, 0.2 mmol) in EtOH (50 mL) was stirred underan atmosphere of H₂ (balloon) for 1 h. The mixture was filtered througha pad of Celite and the filtrate was concentrated under reduced pressureto give 11-(tetrahydro-2H-pyran-4-yl)undecanoic acid (200 mg, 99%) as asolid. LC/MS: mass calcd. for C₁₆H₃₀O₃: 270, found: 269 [M−H]⁻.

Step 3: Synthesis ofN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-11-(tetrahydro-2H-pyran-4-yl)undecanamide

Prepared in a manner similar toN-[(2S,3S,4R)-3,4-dihydroxy-1-{[(2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy}octadecan-2-yl]-11-(3-methyloxetan-3-yl)undecanamide,purified by column chromatography on silica gel (DCM/MeOH 5:1) andpreparative-HPLC to giveN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-11-(tetrahydro-2H-pyran-4-yl)undecanamide(9.6 mg, 8.7%) as a solid. LC/MS: mass calcd. for C₄₀H₇₇NO₁₀: 731.55,found: 732.65 [M+H]⁺; ¹H NMR (400 MHz, CD₃OD) δ 4.21 (d, J=5.7 Hz, 1H),3.56-3.91 (m, 10H), 3.51-3.55 (m, 2H), 3.41-3.44 (m, 2H), 2.22-2.23 (m,2H), 1.65-1.70 (m, 5H), 1.23-1.46 (m, 44H), 0.87-0.97 (m, 3H).

Synthesis of tert-butyl4-(11-(((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)amino)-11-oxoundecyl)piperidine-1-carboxylate

Step 1: Synthesis of11-(1-(tert-butoxycarbonyl)piperidin-4-yl)undec-10-enoic Acid

Prepared in a manner similar to 11-(3-methyloxetan-3-yl)undec-10-enoicacid and purified by column chromatography on silica gel (PE/EtOAc 1:1)to give 11-(1-(tert-butoxycarbonyl)piperidin-4-yl)undec-10-enoic acid(280 mg, 76%) as a solid. LC/MS: mass calcd. for C₂₁H₃₇NO₄: 367, found:366 [M−H]⁻.

Step 2: Synthesis of11-(1-(tert-butoxycarbonyl)piperidin-4-yl)undecanoic Acid

A mixture of 11-(0-(tert-butoxycarbonyl)piperidin-4-yl)undec-10-enoicacid (280 mg, 0.76 mmol) and PtO₂ (40 mg, 0.2 mmol) in EtOH (30 mL) wasstirred under an atmosphere of 112 (balloon) for 1 h. The mixture wasfiltered through a pad of Celite and concentrated under reduced pressureto give 11-(1-(tert-butoxycarbonyl)piperidin-4-yl)undecanoic acid (280mg, 99%) as a solid. LC/MS: mass calcd. for C₂₁H₃₉NO₄: 369, found: 368[M−H]⁻.

Step 3: Synthesis of tert-butyl4-(11-(((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)amino)-11-oxoundecyl)piperidine-1-carboxylate

Prepared in a manner similar toN-[(2S,3S,4R)-3,4-dihydroxy-1-{[(2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy}octadecan-2-yl]-11-(3-methyloxetan-3-yl)undecanamide,purified by column chromatography on silica gel (DCM/MeOH 5:1) andpreparative-HPLC to give tert-butyl4-(11-(((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)amino)-11-oxoundecyl)piperidine-1-carboxylate(13.6 mg, 6%) as a solid. LC/MS: mass calcd. for C₄₅H₈₆N₂O₁₁: 830.62,found: 831.65 [M+H]⁺; ¹H NMR (400 MHz, CD₃OD) δ 4.21 (d, J=5.7 Hz, 1H),4.01-4.07 (m, 2H), 3.56-3.91 (m, 10H), 2.65-2.71 (m, 2H), 2.17-2.19 (m,2H), 1.63-1.70 (m, 6H), 1.54 (s, 9H), 1.33-1.46 (m, 41H), 1.01-1.09 (m,2H), 0.87-0.97 (m, 3H).

Synthesis ofN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-11-(piperidin-4-yl)undecanamide

To a mixture of tert-butyl4-(11-(((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)amino)-11-oxoundecyl)piperidine-1-carboxylate(100 mg, 0.14 mmol) in EtOAc (3 mL) was added 2N HCl in EtOAc (3 mL).The mixture was stirred at rt for 1 h, then concentrated under reducedpressure and the residue was purified by preparative-HPLC to give togiveN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-11-(piperidin-4-yl)undecanamide(10.2 mg, 12.2%) as a solid. LC/MS: mass calcd. for C₄₀H₇₈N₂O₉: 730.57,found: 731.60 [M+H]⁺; ¹H NMR (400 MHz, CD₃OD) δ 4.21 (d, J=5.7 Hz, 1H),3.46-3.91 (m, 10H), 2.89-2.99 (m, 2H), 2.35-2.41 (m, 2H), 1.90-1.97 (m,2H), 1.63-1.70 (m, 5H), 1.33-1.46 (m, 4511), 0.87-0.97 (m, 31).

Synthesis of tert-butyl3-(11-(((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)amino)-11-oxoundecyl)azetidine-1-carboxylate

Step 1: Synthesis of11-(1-(tert-butoxycarbonyl)azetidin-3-yl)undec-10-enoic Acid

Prepared in a manner similar to 11-(3-methyloxetan-3-yl)undec-10-enoicacid and purified by column chromatography on silica gel (PE/EtOAc 1:1)to give 11-(1-(tert-butoxycarbonyl)azetidin-3-yl)undec-10-enoic acid(180 mg, 33%) as a solid. LC/MS: mass calcd. for C₁₉H₃₃NO₄: 339, found:338 [M−H]⁻.

Step 2: Synthesis of 11-[1-(tert-butoxycarbonyl)azetidin-3-yl]undecanoicAcid

A mixture of 11-(1-(tert-butoxycarbonyl)azetidin-3-yl)undec-10-enoicacid (180 mg, 0.53 mmol) and PtO₂ (40 mg, 0.2 mmol) in EtOH (50 mL) wasstirred under an atmosphere of H₂ (balloon) for 1 h. The mixture wasfiltered through a pad of Celite and the filtrate was concentrated underreduced pressure to give11-[1-(tert-butoxycarbonyl)azetidin-3-yl]undecanoic acid (180 mg, 99%)as a solid. LC/MS: mass calcd. for C₁₉H₃₅NO₄: 341, found: 340 [M−H]⁻.

Step 3: Synthesis of tert-butyl3-(11-(((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)amino)-11-oxoundecyl)azetidine-1-carboxylate

Prepared in a manner similar toN-[(2S,3S,4R)-3,4-dihydroxy-1-{[(2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy}octadecan-2-yl]-11-(3-methyloxetan-3-yl)undecanamide,purified by column chromatography on silica gel (DCM/MeOH 5:1) andpreparative-HPLC to give tert-butyl3-(11-(((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)amino)-11-oxoundecyl)azetidine-1-carboxylate(9.4 mg, 6.2%) as a solid. LC/MS: mass calcd. for C₄₃H₈₂N₂O₁₁: 802.59,found: 803.65 [M+H]⁺; ¹H NMR (400 MHz, CD₃OD) δ 4.21 (d, J=5.7 Hz, 1H),3.98-4.05 (m, 2H), 3.46-3.81 (m, 12H), 2.41-2.53 (m, 1H), 2.18-2.25 (m,2H), 1.63-1.70 (m, 6H), 1.50 (s, 9H), 1.33-1.46 (m, 39H), 0.87-0.97 (m,3H).

Synthesis of11-(azetidin-3-yl)-N-((2S,3S,4R)-3,4-dihydroxy-1-((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)undecanamide

To a mixture of tert-butyl3-(11-(((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)amino)-11-oxoundecyl)azetidine-1-carboxylate(100 mg, 0.14 mmol) in EtOAc (3 mL) was added 2N HCl in EtOAc (3 mL).The mixture was stirred at rt for 1 h, then concentrated under reducedpressure and the residue was purified by preparative-HPLC to give11-(azetidin-3-yl)-N-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)undecanamide(7.6 mg, 8.6%) as a solid. LC/MS: mass calcd. for C₃₈H₇₄N₂O₉: 702.54,found: 703.60 [M+H]⁺; ¹H NMR (400 MHz, CD₃OD) δ 4.11-4.21 (m, 2H),3.46-3.93 (m, 10H), 2.80-2.91 (m, 2H), 2.35-2.41 (m, 1H), 2.25-2.29 (m,1H), 1.63-1.70 (m, 5H), 1.33-1.46 (m, 42H), 0.87-0.97 (m, 3H).

Synthesis ofN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-11-(1,1-dioxidotetrahydro-2H-thiopyran-4-yl)undecanamide

Step 1: Synthesis of11-(1,1-dioxidotetrahydro-2H-thiopyran-4-yl)undec-10-enoic Acid

Prepared in a manner similar to 11-(3-methyloxetan-3-yl)undec-10-enoicacid and purified by column chromatography on silica gel (PE/EtOAc 1:1)to give 11-(1,1-dioxidotetrahydro-2H-thiopyran-4-yl)undec-10-enoic acid(100 mg, 51%) as a solid. LC/MS: mass calcd. for CH₁₆H₂₈O₄S: 316, found:315 [M−H]⁻.

Step 2: Synthesis of11-(1,1-dioxidotetrahydro-2H-thiopyran-4-yl)undecanoic Acid

A mixture of 11-(0,1-dioxidotetrahydro-2H-thiopyran-4-yl)undec-10-enoicacid (80 mg, 0.3 mmol) and PtO₂ (20 mg, 0.1 mmol) in EtOH (30 mL) wasstirred under an atmosphere of H₂ (balloon) for 1 h. The mixture wasfiltered through a pad of Celite and the filtrate was concentrated underreduced pressure to give11-(1,1-dioxidotetrahydro-2H-thiopyran-4-yl)undecanoic acid (80 mg, 80%)as a solid. LC/MS: mass calcd. for C₁₆H₃₀O₄S: 318, found: 317 [M−H]⁻.

Step 3: Synthesis ofN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-1-(1,1-dioxidotetrahydro-2H-thiopyran-4-yl)undecanamide

Prepared in a manner similar toN-[(2S,3S,4R)-3,4-dihydroxy-1-{[(2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy}octadecan-2-yl]-11-(3-methyloxetan-3-yl)undecanamide,purified by column chromatography on silica gel (DCM/MeOH 5:1) andpreparative-HPLC to giveN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-11-(1,1-dioxidotetrahydro-2H-thiopyran-4-yl)undecanamide(20.7 mg, 21%) as a solid. LC/MS: mass calcd. for C₄₀H₇₇NO₁₁S: 779.52,found: 780.50 [M+H]⁺; ¹H NMR (300 MHz, DMSO-d₆) δ 4.65 (s, 1H),3.33-3.93 (m, 12H), 2.93-3.13 (m, 4H), 2.93 (d, J=13.5 Hz, 2H), 2.04 (t,J=7.2 Hz, 2H), 1.94 (d, J=10.3 Hz, 2H), 1.38-1.53 (m, 8H), 1.10-1.32 (m,43H), 0.87-0.97 (m, 3H).

Synthesis of tert-butyl3-(11-(((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)amino)-11-oxoundecyl)pyrrolidine-1-carboxylate

Step 1: Synthesis of11-(1-(tert-butoxycarbonyl)pyrrolidin-3-yl)undec-10-enoic Acid

Prepared in a manner similar to 11-(3-methyloxetan-3-yl)undec-10-enoicacid and purified by column chromatography on silica gel (PE/EtOAc 1:1)to give 11-(1-(tert-butoxycarbonyl)pyrrolidin-3-yl)undec-10-enoic acid(150 mg, 42%) as a solid. LC/MS: mass calcd. for C₂₀H₃₅NO₄: 353, found:352 [M−H]⁻.

Step 2: Synthesis of11-(1-(tert-butoxycarbonyl)pyrrolidin-3-yl)undecanoic Acid

A mixture of 11-[1-(tert-butoxycarbonyl)pyrrolidin-3-yl]undec-10-enoicacid (150 mg, 0.3 mmol) and PtO₂ (20 mg, 0.1 mmol) in EtOH (50 mL) wasstirred under an atmosphere of H₂ (balloon) for 1 h. The reactionmixture was filtered through a pad of Celite and the filtrate wasconcentrated under reduced pressure to afford11-(1-(tert-butoxycarbonyl)pyrrolidin-3-yl)undecanoic acid (140 mg, 93%)as a solid. LC/MS: mass calcd. for C₂₀H₃₇NO₄. 355, found: 354 [M−H]⁻.

Step 3: Synthesis of tert-butyl3-(11-(((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)amino)-11-oxoundecyl)pyrrolidine-1-carboxylate

Prepared in a manner similar toN-[(2S,3S,4R)-3,4-dihydroxy-1-{[(2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy}octadecan-2-yl]-11-(3-methyloxetan-3-yl)undecanamide,purified by column chromatography on silica gel (DCM/MeOH 5:1) andpreparative-HPLC to give tert-butyl3-(11-(((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)amino)-11-oxoundecyl)pyrrolidine-1-carboxylate(6.1 mg, 3.6%) as a solid. LC/MS: mass calcd. for C₄₄H₈₄N₂O₁₁: 816.61,found: 839.55 [M+Na]⁺, ¹H NMR (300 MHz, CD₃OD) δ 4.18 (d, J=5.9 Hz, 1H),3.35-3.92 (m, 11H), 3.15-3.20 (m, 2H), 2.82 (s, 1H), 1.91-2.34 (m, 4H),1.02-1.77 (m, 54H), 0.90 (t, J=6.6 Hz, 3H).

Synthesis ofN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-11-(pyrrolidin-3-yl)undecanamide

To a mixture of tert-butyl3-(11-(((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)amino)-11-oxoundecyl)pyrrolidine-1-carboxylate(60 mg, 0.07 mmol) in EtOAc (3 mL) was added 2N HCl in EtOAc (3 mL). Themixture was stirred at rt for 1 h, then concentrated under reducedpressure and purified by preparative-HPLC to giveN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-11-(pyrrolidin-3-yl)undecanamide(15.6 mg, 28%) as an oil. LC/MS: mass calcd. for C₃₉H₇₆N₂O₉: 716.56,found: 717.50 [M+H]⁺; ¹H NMR (300 MHz, CD₃OD) δ 4.21 (d, J=5.7 Hz, 1H),3.51-3.91 (m, 14H), 3.12-3.21 (m, 4H), 2.74-2.79 (m, 1H), 2.15-2.30 (m,3H), 1.14-1.61 (m, 42H), 0.87-0.97 (m, 3H).

Synthesis ofN-[(2S,3S,4R)-3,4-dihydroxy-1-{[3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy}octadecan-2-yl]-11-{2-oxa-6-azaspiro[3.3]heptan-6-yl}undecanamide

Step 1: Synthesis of ter-butyl11-(2-oxa-6-azaspiro[3.3]heptan-6-yl)undecanoate

To a mixture of 2-oxa-6-azaspiro[3.3]heptane (200 mg, 2.0 mmol) in DMF(10 mL) was added tert-butyl 11-bromoundecanoate (642 mg, 2.0 mmol) andK₂CO₃ (552 mg, 4.0 mmol). The mixture was heated to 100° C., and stirredfor 5 h, then diluted with H₂O (20 mL) and extracted with EtOAc (3×30mL). The combined organic layers were washed with brine (50 mL), driedover anhydrous Na₂SO₄, filtered, and the filtrate was concentrated underreduced pressure. The residue was purified by column chromatography onsilica gel (PE/EtOAc 1:1) to give tert-butyl11-(2-oxa-6-azaspiro[3.3]heptan-6-yl)undecanoate (339 mg, 50%) as asolid. LC/MS: mass calcd. for C₂₀H₃₇NO₃: 339, found: 340 [M+H]⁺.

Step 2: Synthesis of 11-(2-oxa-6-azaspiro[3.3]heptan-6-yl)undecanoicAcid

A mixture of tert-butyl 11-(2-oxa-6-azaspiro[3.3]heptan-6-yl)undecanoate(220 mg, 0.6 mmol), DCM (4 mL) and TFA (2 mL) was stirred at rt for 2 h,then concentrated under reduced pressure to give11-(2-oxa-6-azaspiro[3.3]heptan-6-yl)undecanoic acid (180 mg, 98%) as asolid.

Step 3: Synthesis ofN-[(2S,3S,4R)-3,4-dihydroxy-1-{[3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy}octadecan-2-yl]-11-{2-oxa-6-azaspiro[3.3]heptan-6-yl}undecanamide

Prepared in a manner similar toN-[(2S,3S,4R)-3,4-dihydroxy-1-{[(2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy)}octadecan-2-yl]-11-(3-methyloxetan-3-yl)undecanamide,purified by column chromatography on silica gel (DCM/MeOH 5:1) andpreparative-HPLC to giveN-[(2S,3S,4R)-3,4-dihydroxy-1-{[3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy}octadecan-2-yl]-11-{2-oxa-6-azaspiro[3.3]heptan-6-yl}undecanamide(8.5 mg, 6.8%) as a solid. LC/MS: mass calcd. for C₄₀H₇₆N₂O₁₀: 744.55,found: 745.55 [M+H]⁺; ¹H NMR (300 MHz, CD₃OD) δ 4.75 (s, 4H), 4.14-4.24(m, 1H), 3.56-3.87 (m, 15H), 2.68-2.79 (m, 1H), 2.15-2.25 (m, 2H),1.49-1.66 (m, 5H), 1.33-1.46 (m, 47H), 0.87-0.97 (m, 3H).

Synthesis ofN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-11-morpholinoundecanamide

Step 1: Synthesis of tert-butyl 11-morpholinoundecanoate

Prepared in a manner similar to tert-butyl11-(2-oxa-6-azaspiro[3.3]heptan-6-yl)undecanoate and purified by columnchromatography on silica gel (PE/EtOAc 1:1) to give tert-butyl11-morpholinoundecanoate (450 mg, 88%) as a solid. LC/MS: mass calcd.for C₁₉H₃₇NO₃: 327, found: 328 [M+H]⁺.

Step 2: Synthesis of 11-morpholinoundecanoic Acid

To a mixture of tert-butyl 11-(morpholin-4-yl)undecanoate (450 mg, 1.3mmol) and EtOAc (5 mL) was added 2N HCl in EtOAc (5 mL). The mixture wasstirred at rt for 1 h, then concentrated under reduced pressure to give11-morpholinoundecanoic acid (300 mg, 77%) as a solid.

Step 3: Synthesis ofN-((2S,3S,4R)-3,4-bis(benzyloxy)-1-(((2S,3R,4S,5S,6R)-3,4,5-tris(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-11-morpholinoundecanamide

Prepared in a manner similar toN-[(2S,3S,4R)-3,4-dihydroxy-1-{[(2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy}octadecan-2-yl]-11-(3-methyloxetan-3-yl)undecanamide.Purified by column chromatography on silica gel (PE/EtOAc 2:1) to giveN-((2S,3S,4R)-3,4-bis(benzyloxy)-1-(((2S,3R,4S,5S,6R)-3,4,5-tris(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-11-morpholinoundecanamide(70 mg, 28%) as a solid.

Step 4: Synthesis ofN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-11-morpholinoundecanamide

A mixture ofN-((2S,3S,4R)-3,4-bis(benzyloxy)-1-(((2S,3R,4S,5S,6R)-3,4,5-tris(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-11-morpholinoundecanamide(70 mg, 0.055 mmol) and Pd(OH)₂/C (20 mg) in DCM (5 mL) and EtOH (5 mL)was stirred under an atmosphere of H₂ (balloon) for 16 h. The mixturewas filtered through a pad of Celite, the filtrate was concentratedunder reduced pressure and the residue was purified by preparative-HPLCto giveN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-11-morpholinoundecanamide(9.1 mg, 23%) as a solid. LC/MS: mass calcd. for C₃₉H₇₆N₂O₁₀: 732.55,found: 733.50 [M+H]⁺. ¹H NMR (300 MHz, CD₃OD) 4.21 (d, J=5.7 Hz, 1H),4.02-4.09 (m, 2H), 3.63-3.87 (9H), 3.46-3.58 (m, 4H), 3.11-3.16 (4H),2.22 (t, J=7.4 Hz, 2H), 1.33-1.70 (m, 43H), 0.87-0.97 (m, 3H).

Synthesis ofN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-11-(piperidin-1-yl)undecanamide

Step 1: Synthesis of tert-butyl 11-(piperidin-1-yl)undecanoate

Prepared in a manner similar to tert-butyl11-(2-oxa-6-azaspiro[3.3]heptan-6-yl)undecanoate and purified by columnchromatography on silica gel (PE/EtOAc 2:1) to give tert-butyl11-(piperidin-1-yl)undecanoate (240 mg, 47%) as a solid. LC/MS: masscalcd. for C₂₀H₃₉NO₂: 325, found: 326 [M+H]⁺.

Step 2: Synthesis of 11-(piperidin-1-yl)undecanoic Acid

To a mixture of tert-butyl 11-(piperidin-1-yl)undecanoate (180 mg, 0.5mmol) and EtOAc (3 mL) was added 2N HCl in EtOAc (3 mL). The mixture wasstirred at rt for 1 h, then concentrated under reduced pressure to give11-(piperidin-1-yl)undecanoic acid (140 mg, 94%) as a solid.

Step 3: Synthesis ofN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-11-(piperidin-1-yl)undecanamide

Prepared in a manner similar toN-[(2S,3S,4R)-3,4-dihydroxy-1-{[(2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy}octadecan-2-yl]-11-(3-methyloxetan-3-yl)undecanamide,purified by column chromatography on silica gel (DCM/MeOH 5:1) andpreparative-HPLC to giveN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-11-(piperidin-1-yl)undecanamide(32 mg, 26%) as an oil. LC/MS: mass calcd. for C₄₀H₇₈N₂O₉: 730.57,found: 731.55 [M+H]⁺; ¹H NMR (400 MHz, CD₃OD) δ 4.23 (q, J=4.9 Hz, 1H),3.68-3.92 (m, 7H), 3.50-3.61 (m, 5H), 3.07-3.09 (m, 2H), 2.89-2.97 (m,2H), 2.18-2.30 (m, 2H), 1.96-1.99 (m, 1H), 1.50-1.87 (m, 11H), 1.30-1.39(m, 36H), 0.87-0.97 (m, 3H).

Synthesis ofN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-11-(pyrrolidin-1-yl)undecanamide

Step 1: Synthesis of tert-butyl 11-(pyrrolidin-1-yl)undecanoate

Prepared in a manner similar to tert-butyl11-(2-oxa-6-azaspiro[3.3]heptan-6-yl)undecanoate and purified by columnchromatography on silica gel (PE/EtOAc 1:1) to give tert-butyl11-(pyrrolidin-1-yl)undecanoate (310 mg, 64%) as a solid. LC/MS: masscalcd. for C₁₉H₃₇NO₂: 311, found: 312 [M+H]⁺.

Step 2: Synthesis of 11-(pyrrolidin-1-yl)undecanoic Acid

To a mixture of tert-butyl 11-(pyrrolidin-1-yl)undecanoate (310 mg, 0.99mmol) and EtOAc (3 mL) was added 2N HCl in EtOAc (3 mL). The mixture wasstirred at rt for 1 h, then concentrated under reduced pressure to give11-(pyrrolidin-1-yl)undecanoic acid (240 mg, 94%) as a solid.

Step 3: Synthesis ofN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H1-pyran-2-yl)oxy)octadecan-2-yl)-11-(pyrrolidin-1-yl)undecanamide

Prepared in a manner similar toN-[(2S,3S,4R)-3,4-dihydroxy-1-{[(2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy}octadecan-2-yl]-11-(3-methyloxetan-3-yl)undecanamide,purified by column chromatography on silica gel (DCM/MeOH 5:1) andpreparative-HPLC to giveN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-11-(pyrrolidin-1-yl)undecanamide(15 mg, 12.4%) as an oil LC/MS: mass calcd. for C₃₉H₇₆N₂O₉: 716.56,found: 717.50 [M+H]⁺; ¹H NMR (300 MHz, CD₃OD) δ 4.21 (d, J=5.7 Hz, 1H),3.56-3.91 (m, 10H), 3.13-3.18 (m, 2H), 3.02-3.11 (m, 2H), 2.13-2.23 (m,4H), 1.94-2.06 (m, 2H), 1.54-1.75 (m, 7H), 1.27-1.39 (m, 37H), 0.87-4.97(m, 3H).

Synthesis ofN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-11-thiomorpholinoundecanamide

Step 1: Synthesis of tert-butyl 11-thiomorpholinoundecanoate

Prepared in a manner similar to tert-butyl111-(2-oxa-6-azaspiro[3.3]heptan-6-yl)undecanoate and purified by columnchromatography on silica gel (PE/EtOAc 2:1) to give tert-butyl11-thiomorpholinoundecanoate (340 mg, 64%) as a solid. LC/MS: masscalcd. for C₁₉H₃₇NO₂S: 343, found: 344 [M+H]⁺.

Step 2: Synthesis of 11-thiomorpholinoundecanoic Acid

To a mixture of tert-butyl 11-thiomorpholinoundecanoate (400 mg, 1.2mmol) and EtOAc (5 mL) was added 2N HCl in EtOAc (5 mL). The mixture wasstirred at rt for 1 h at rt, then concentrated under reduced pressure togive 11-thiomorpholinoundecanoic acid (318 mg, 95%) as a solid.

Step 3: Synthesis ofN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-11-thiomorpholinoundecanamide

Prepared in a manner similar toN-[(2S,3S,4R)-3,4-dihydroxy-1-{[(2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy}octadecan-2-yl]-11-(3-methyloxetan-3-yl)undecanamide,purified by column chromatography on silica gel (DCM/MeOH 5:1) andpreparative-HPLC to giveN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-11-thiomorpholinoundecanamide(17.1 mg, 13.6%) as an oil. LC/MS: mass calcd. for C₃₉H₇₆N₂O₉S: 748.53,found: 749.45 [M+H]⁺; ¹H NMR (400 MHz, CD₃OD) δ 4.23 (q, J=4.9 Hz, 1H),3.65-3.96 (m, 10H), 3.03-3.29 (m, 6H), 287-2.91 (m, 3H), 2.19-2.32 (m,2H), 1.52-1.79 (m, 6H), 1.30-1.39 (m, 39H), 0.89-0.96 (m, 3H).

Synthesis ofN-[(2S,3S,4R)-3,4-dihydroxy-1-{[3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy}octadecan-2-yl]-11-[(1S,5S)-6,6-dimethylbicyclo[3.1.1]heptan-2-yl]undecanamide

Step 1: Synthesis of11-((1R,5S)-6,6-dimethylbicyclo[3.1.1]hept-2-en-2-yl)undec-10-enoic Acid

Prepared in a manner similar to 11-(3-methyloxetan-3-yl)undec-10-enoicacid and purified by column chromatography on silica gel (PE/EtOAc 1:1)to give11-((1R,5S)-6,6-dimethylbicyclo[3.1.1]hept-2-en-2-yl)undec-10-enoic acid(110 mg, 54%) as a solid. LC/MS: mass calcd. for C₂₀H₃₂O₂: 304, found:303 [M−H]⁻.

Step 2: Synthesis of11-((1S,5S)-6,6-dimethylbicyclo[3.1.1]heptan-2-yl)undecanoic Acid

A mixture of11-((1R,5S)-6,6-dimethylbicyclo[3.1.1]hept-2-en-2-yl)undec-10-enoic acid(100 mg, 0.3 mmol) and PtO₂ (20 mg, 0.1 mmol) in EtOH (50 mL) wasstirred under an atmosphere of H₂ (balloon) for 1 h. The mixture wasfiltered through a pad of Celite and the filtrate was concentrated underreduced pressure to give11-((1S,5S)-6,6-dimethylbicyclo[3.1.1]heptan-2-yl)undecanoic acid (80mg, 79%) as a solid. LC/MS: mass calcd. for C₂₀H₃₆O₂: 308, found: 307[M−H]⁻.

Step 3: Synthesis ofN-[(2S,3S,4R)-3,4-dihydroxy-1-{[3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy}octadecan-2-yl]-11-[(1S,5S)-6,6-dimethylbicyclo[3.1.1]heptan-2-yl]undecanamide

Prepared in a manner similar toN-[(2S,3S,4R)-3,4-dihydroxy-1-{[(2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy}octadecan-2-yl]-11-(3-methyloxetan-3-yl)undecanamide,purified by column chromatography on silica gel (DCM/MeOH 5:1) andpreparative-HPLC to giveN-[(2S,3S,4R)-3,4-dihydroxy-1-{[3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy}octadecan-2-yl]-11-[(1S,5S)-6,6-dimethylbicyclo[3.1.1]heptan-2-yl]undecanamide(14.3 mg, 10.7%) as a solid. LC/MS: mass calcd. for C₄₄H₈₃NO₉: 769.61,found: 792.65 [M+Na]⁺; ¹H NMR (300 MHz, CD₃OD) δ 4.17 (dd, J=6.7, 4.1Hz, 1H), 3.51-3.92 (m, 10H), 3.17-3.26 (m, 3H), 2.31-2.38 (m, 1H),2.18-2.22 (m, 2H), 1.85-1.97 (m, 4H), 1.52-1.62 (m, 5H), 1.22-1.39 (m,44H), 1.19 (s, 3H), 0.87-0.97 (m, 3H).

Synthesis ofN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-11-(1,1-dioxidothiomorpholino)undecanamide

Step 1: Synthesis of tert-butyl11-(2-oxa-6-azaspiro[3.3]heptan-6-yl)undecanoate

Prepared in a manner similar to tert-butyl11-(2-oxa-6-azaspiro[3.3]heptan-6-yl)undecanoate and purified by columnchromatography on silica gel (PE/EtOAc 1:1) to give tert-butyl11-(1,1-dioxidothiomorpholino)undecanoate (320 mg, 55%) as a solidLC/MS: mass calcd. for C₁₉H₃₇NO₄S: 375, found: 376 [M+H]⁺.

Step 2: Synthesis of 11-(1,1-dioxidothiomorpholino)undecanoic Acid

To a mixture of tert-butyl 11-(1,1-dioxidothiomorpholino)undecanoate(300 mg, 0.79 mmol) and EtOAc (3 mL) was added 2N HCl in EtOAc (3 mL).The mixture was stirred at rt for 1 h, then concentrated under reducedpressure to give 11-(1,1-dioxidothiomorpholino)undecanoic acid (200 mg,78%) as a solid. LC/MS: mass calcd. for C₁₅H₂₉NO₄S: 319, found: 318[M−H]⁻.

Step 3: Synthesis ofN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-11-(1,1-dioxidothiomorpholino)undecanamide

Prepared in a manner similar toN-[(2S,3S,4R)-3,4-dihydroxy-1-{[(2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy}octadecan-2-yl]-11-(3-methyloxetan-3-yl)undecanamide,purified by column chromatography on silica gel (DCM/MeOH 5:1) andpreparative-HPLC to giveN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-11-(1,1-dioxidothiomorpholino)undecanamide(17.3 mg, 16.2%) as a solid. LC/MS: mass calcd. for C₃₉H₇₆N₂O₁₁S:780.52, found: 704.50 [M+H]⁺; ¹H NMR (300 MHz, CD₃OD) δ 4.19 (q, J=4.8Hz, 1H), 3.93-3.51 (m, 10H), 3.08 (q, J=4.6 Hz, 4H), 2.97 (dd, J=6.8,3.4 Hz, 4H), 2.56-2.45 (m, 2H), 2.22 (t, J=7.5 Hz, 2H), 1.11-1.61 (m,42H), 0.95-0.85 (m, 3H).

Synthesis ofN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-11-(isopropylsulfonyl)undecanamide

Step 1: Synthesis of tert-butyl 11-(isopropylthio)undecanoate

To a mixture of 2-propanethiol (3.5 g, 46.6 mmol) and DMF (30 mL) wasadded tert-butyl 11-bromoundecanoate (1.0 g, 3.1 mmol) and Cs₂CO₃ (2.5g, 7.7 mmol). The mixture was stirred at rt 5 h, then diluted with H₂O(60 mL) and extracted with EtOAc (3×50 mL). The combined organic layerswere washed with brine (60 mL), dried over anhydrous Na₂SO₄, filtered,and the filtrate was concentrated under reduced pressure. The residuewas purified by column chromatography on silica gel (PE/EtOAc 2:1) togive tert-butyl 11-(isopropylthio)undecanoate (900 mg, 91%) as a solid.

Step 2: Synthesis of Tert-butyl 11-(isopropylsulfonyl)undecanoate

To a mixture of tert-butyl 11-(isopropylthio)undecanoate (800 mg, 2.5mmol) in DCM (20 mL) was added m-CPBA (870 mg, 5.0 mmol). The mixturewas stirred at rt overnight, then diluted with H₂O (50 mL) and extractwith EtOAc (3×50 mL). The combined organic layers were washed with brine(50 mL), dried over anhydrous Na₂SO₄, filtered and the filtrate wasconcentrated under reduced pressure. The residue was purified by columnchromatography on silica gel (PE/EtOAc 2:1) to give tert-butyl11-(isopropylsulfonyl)undecanoate (600 mg, 68%) as a solid.

Step 3: Synthesis of 11-(isopropylsulfonyl)undecanoic Acid

A mixture of tert-butyl 11-(isopropylsulfonyl)undecanoate (300 mg, 0.86mmol) in DCM (4 mL) and TFA (2 mL) was stirred at rt for 2 h, thenconcentrated under reduced pressure to give11-(isopropylsulfonyl)undecanoic acid (200 mg, 79%) as a solid. LC/MS:mass calcd. for C₁₄H₂₈O₄S: 292, found: 291 [M−H]⁻.

Step 4: Synthesis ofN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-11-(isopropylsulfonyl)undecanamide

Prepared in a manner similar toN-[(2S,3S,4R)-3,4-dihydroxy-1-{[(2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy}octadecan-2-yl]-11-(3-methyloxetan-3-yl)undecanamide,purified by column chromatography on silica gel (DCM/MeOH 5:1) andpreparative-HPLC to giveN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-11-(isopropylsulfonyl)undecanamide(31.3 mg, 28%) as a solid. LC/MS: mass calcd. for C₃₈H₇₅NO₁₁S: 753.51,found: 754.50 [M+H]⁺; ¹H NMR (400 MHz, CD₃OD) δ 4.89 (d, J=3.7 Hz, 1H),4.22 (q, J=4.8 Hz, 1H), 3.52-3.94 (m, 10H), 3.22-3.28 (m, 1H), 3.02-3.11(m, 2H), 2.24 (t, J=7.5 Hz, 2H), 1.88-1.75 (m, 2H), 1.30-1.66 (m, 46H),0.96-0.88 (m, 3H).

Synthesis ofN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-21-pyran-2-yl)oxy)octadecan-2-yl)-10-((1,1-dioxidotetrahydro-2H-thiopyran-4-yl)oxy)decanamide

Step 1: Synthesis of methyl10-((1,1-dioxidotetrahydro-2H-thiopyran-4-yl)oxy)decanoate

To a mixture of 4-hydroxy tetrahydro-2H-thiopyran 1,1-dioxide (200 mg,1.3 mmol) and THF (10 mL) at 0° C. under an atmosphere of N₂ was added 2M NaHMDS (0.65 mL, 1.3 mmol). The mixture was warmed to rt and stirredfor 1 h, then a solution of methyl 10-bromodecanoate (235 mg, 0.9 mmol)in THF (1.5 mL) was added. The mixture was stirred at rt overnight, thendiluted with H₂O (20 mL) and extracted with EtOAc (3×30 mL). Thecombined organic layers were washed with brine (30 mL), dried overanhydrous Na₂SO₄, filtered, and the filtrate was concentrated underreduced pressure. The residue was purified by column chromatography onsilica gel (PE/EtOAc 1:1) to give methyl10-((1,1-dioxidotetrahydro-2H-thiopyran-4-yl)oxy)decanoate (80 mg, 27%)as a solid. LC/MS: mass calcd. for C₁₆H₃₀O₅S: 334, found: 335 [M+H]⁺.

Step 2: Synthesis of10-((1,1-dioxidotetrahydro-2H-thiopyran-4-yl)oxy)decanoic Acid

To a mixture of methyl10-((1,1-dioxidotetrahydro-2H-thiopyran-4-yl)oxy)decanoate (200 mg, 0.6mmol) in MeOH (5 mL), THF (5 mL) and H₂O (5 mL) was added LiOH (42 mg,1.8 mmol). The mixture was stirred at rt for 2 h, then acidified pH˜3with 1N HCl and extracted with EtOAc (3×30 mL). The combined organiclayers were washed with brine (30 mL), dried over anhydrous Na₂SO₄,filtered, and the filtrate was concentrated under reduced pressure. Theresidue was purified column chromatography on by silica gel (DCM/MeOH10:1) to give 10-((1,1-dioxidotetrahydro-2H-thiopyran-4-yl)oxy)decanoicacid (66 mg, 53%) as a solid. LC/MS: mass calcd. for C₁₅H₂₈O₅S: 320,found: 319 [M−H]⁻.

Step 3: Synthesis ofN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-10-((1,1-dioxidotetrahydro-2H1-thiopyran-4-yl)oxy)decanamide

Prepared in a manner similar toN-[(2S,3S,4R)-3,4-dihydroxy-1-{[(2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy}octadecan-2-yl]-11-(3-methyloxetan-3-yl)undecanamide,purified by column chromatography on silica gel (DCM/MeOH 5:1) andpreparative-HPLC to giveN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-10-((1,1-dioxidotetrahydro-2H-thiopyran-4-yl)oxy)decanamide(22.2 mg, 14.5%) as an oil. LC/MS: mass calcd. for C₃₉H₇₅NO₁₂S: 781.50,found: 782.45 [M+H]⁺; ¹H NMR (400 MHz, CD₃OD) δ 4.20 (dd, J=6.4, 4.5 Hz,1H), 3.52-3.89 (m, 10H), 3.46-3.48 (m, 2H), 3.15-3.30 (m, 2H), 2.91-2.95(m, 2H), 2.08-2.28 (m, 5H), 1.53-1.63 (m, 6H), 1.27-1.39 (m, 36H),0.94-0.86 (m, 3H).

Synthesis ofN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-10-((tetrahydro-2H-pyran-4-yl)oxy)decanamide

Step 1: Synthesis of methyl 10-((tetrahydro-2H-pyran-4-yl)oxy)decanoate

Prepared in a manner similar to methyl10-((1,1-dioxidotetrahydro-2H-thiopyran-4-yl)oxy)decanoate and purifiedby column chromatography on silica gel (PE/EtOAc 2:1) to give methyl10-((tetrahydro-21H-pyran-4-yl)oxy)decanoate (95 mg, 37%) as an oil.

Step 2: Synthesis of 10-((tetrahydro-2H-pyran-4-yl)oxy)decanoic Acid

Prepared in a manner similar to10-((1,1-dioxidotetrahydro-2H-thiopyran-4-yl)oxy)decanoic acid andpurified by column chromatography on silica gel (DCM/MeOH 10:1) to give10-((tetrahydro-2H-pyran-4-yl)oxy)decanoic acid (65 mg, 80%) as a solid.LC/MS: mass calcd. for C₁₅H₂₈O₄: 272, found: 271 [M−H]⁻.

Step 3: Synthesis ofN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-10-((tetrahydro-2H-pyran-4-yl)oxy)decanamide

Prepared in a manner similar toN-[(2,S,3S,4R)-3,4-dihydroxy-1-{[(2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy}octadecan-2-yl]-11-(3-methyloxetan-3-yl)undecanamide,purified by column chromatography on silica gel (DCM/MeOH 5:1) andpreparative-HPLC to giveN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-10-((tetrahydro-2H-pyran-4-yl)oxy)decanamide(12 mg, 13%) as an oil. LC/MS: mass calcd. for C₃₉H₇₅NO₁₁: 733.53,found: 734.50 [M+H]⁺. ¹H NMR (300 MHz, CD₃OD) 4.19 (q, J=4.9 Hz, 1H),3.66-3.90 (m, 9H), 3.37-3.51 (m, 5H), 2.22 (t, J=7.5 Hz, 2H), 1.89 (dd,J=13.0, 3.8 Hz, 2H), 1.47-1.62 (m, 5H), 1.26-1.37 (m, 40H), 0.88-0.91(m, 3H).

Synthesis ofN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-21-pyran-2-yl)oxy)octadecan-2-yl)-9-((tetrahydro-2H1-pyran-4-yl)methoxy)nonanamide

Step 1: Synthesis of ethyl9-((tetrahydro-2H-pyran-4-yl)methoxy)nonanoate

Prepared in a manner similar to methyl10-((1,1-dioxidotetrahydro-2H-thiopyran-4-yl)oxy)decanoate and purifiedby column chromatography on silica gel (PE/EtOAc 2:1) to give ethyl9-((tetrahydro-2H-pyran-4-yl)methoxy)nonanoate (94 mg, 29%) as an oil.

Step 2: Synthesis of 9-((tetrahydro-2H-pyran-4-yl)methoxy)nonanoic acid

Prepared in a manner similar to10-((1,1-dioxidotetrahydro-2H-thiopyran-4-yl)oxy)decanoic acid andpurified by column chromatography on silica gel (DCM/MeOH 10:1) to give9-((tetrahydro-2H-pyran-4-yl)methoxy)nonanoic acid (57 mg, 70%) as asolid. LC/MS: mass calcd. for C₁₅H₂₈O₄: 272, found: 271 [M−H]⁻.

Step 3: Synthesis ofN-((2S,3S,4R)-3,4-bis(benzyloxy)-1-(((2S,3R,4S,5S,6R)-3,4,5-tris(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-9-((tetrahydro-2H-pyran-4-yl)methoxy)nonanamide

Prepared in a manner similar toN-[(2S,3S,4R)-3,4-dihydroxy-1-{[(2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy}octadecan-2-yl]-11-(3-methyloxetan-3-yl)undecanamide,purified by column chromatography on silica gel (PE/EtOAc 5:1) to giveN-((2S,3S,4R)-3,4-bis(benzyloxy)-1-(((2S,3R,4S,5S,6R)-3,4,5-tris(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-9-((tetrahydro-2H-pyran-4-yl)methoxy)nonanamideas a solid. LC/MS: mass calcd. for C₈₁H₁₁₁NO₁₁: 1273.82, found: 1274.50[M+H]⁺.

Step 4: Synthesis ofN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-9-((tetrahydro-2H-pyran-4-yl)methoxy)nonanamide

A mixture ofN-((2S,3S,4R)-3,4-bis(benzyloxy)-1-(((2S,3R,4S,5S,6R)-3,4,5-tris(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-9-((tetrahydro-2H-pyran-4-yl)methoxy)nonanamide(20 mg, 0.013 mmol) and Pd(OH)₂ (40 mg) in EtOH (3 mL) and DCM (3 mL)was stirred under an atmosphere of H₂ (balloon) for 16 h. The mixturewas filtered through a pad of Celite and the filtrate was concentratedunder reduced pressure to giveN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-9-((tetrahydro-2H1-pyran-4-yl)methoxy)nonanamide(6.3 mg, 54%) as a solid. LC/MS: mass calcd. for C₃₉H₇₅NO₁₁: 733.53,found: 734.50 [M+H]⁺; ¹H NMR (300 MHz, CD₃OD) δ 4.22 (d, J=5.8 Hz, 1H),3.56-3.94 (m, 10H), 3.40-3.48 (m, 3H), 3.28-3.30 (m, 3H), 2.24 (t, J=7.5Hz, 2H), 1.54-1.69 (m, 6H), 1.29-1.35 (m, 39H), 0.97-0.87 (m, 3H).

Synthesis ofN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-21-pyran-2-yl)oxy)octadecan-2-yl)-8-(2-(tetrahydro-2H-pyran-4-yl)ethoxy)octanamide

Step 1: Synthesis of ethyl8-(2-(tetrahydro-2H-pyran-4-yl)ethoxy)octanoate

Prepared in a manner similar to methyl10-((1,1-dioxidotetrahydro-2H-thiopyran-4-yl)oxy)decanoate and purifiedby column chromatography on silica gel (PE/EtOAc 2:1) to give ethyl8-(2-(tetrahydro-2H-pyran-4-yl)ethoxy)octanoate (62 mg, 23%) as an oil.

Step 2: Synthesis of 8-(2-(tetrahydro-2H-pyran-4-yl)ethoxy)octanoic Acid

Prepared in a manner similar to10-((1,1-dioxidotetrahydro-2H-thiopyran-4-yl)oxy)decanoic acid andpurified by column chromatography on silica gel (DCM/MeOH 10:1) to give8-(2-(tetrahydro-2H-pyran-4-yl)ethoxy)octanoic acid (47 mg, 86%) as asolid. LC/MS: mass calcd. for C₁₅H₂₈O₄: 272, found: 271 [M−H]⁻.

Step 3: Synthesis ofN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-8-(2-(tetrahydro-2H-pyran-4-yl)ethoxy)octanamide

Prepared in a manner similar toN-[(2S,3S,4R)-3,4-dihydroxy-1-{[(2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy}octadecan-2-yl]-11-(3-methyloxetan-3-yl)undecanamide,purified by column chromatography on silica gel (DCM/MeOH 5:1) andpreparative-HPLC to giveN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-8-(2-(tetrahydro-2H-pyran-4-yl)ethoxy)octanamide(7.4 mg, 8.4%) as an oil. LC/MS: mass calcd. for C₃₉H₇₅NO₁₁: 733.53,found: 734.45 [M+H]⁺; ¹H NMR (300 MHz, CD₃OD) δ 4.56-4.60 (m, 4H),4.17-4.23 (m, 1H), 3.66-3.92 (m, 15H), 3.39-3.57 (m, 4H), 2.22 (t, J=7.5Hz, 2H), 1.48-1.65 (m, 5H), 1.27-1.37 (m, 33H), 0.84-0.95 (m, 3H).

Synthesis ofN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-7-(3-(tetrahydro-2H-pyran-4-yl)propoxy)heptanamide

Step 1: Synthesis of 7-(3-(tetrahydro-2H-pyran-4-yl)propoxy)heptanoicAcid

To a mixture of 3-(tetrahydro-2H-pyran-4-yl)propan-1-ol (1.0 g, 6.9mmol) and DMSO (20 mL) under an atmosphere of N₂ was added KOH (320 mg,5.7 mmol). The mixture was heated to 80° C., and stirred for 1 h, then7-bromo-heptanoic acid (300 mg, 1.4 mmol) was added and the mixturestirred for at 80° C. for a further 2 h. After cooling, the mixture wasacidified pH˜1 with 1N HCl and extracted with EtOAc (30 mL×3). Thecombined organic layers were washed with brine (30 mL), dried overanhydrous Na₂SO₄, filtered and the filtrate was concentrated underreduced pressure. The residue was purified by column chromatography onsilica gel (DCM/MeOH 10:1) to give7-(3-(tetrahydro-2H-pyran-4-yl)propoxy)heptanoic acid (50 mg, 13%) as anoil. LC/MS: mass calcd. for C₁₅H₂₈O₄: 272, found: 271 [M−H]⁻.

Step 2: Synthesis ofN-((2S,3S,4R)-3,4-bis(benzyloxy)-1-(((2S,3R,4S,5S,6R)-3,4,5-tris(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-7-(3-(tetrahydro-2H-pyran-4-yl)propoxy)heptanamide

Prepared in a manner similar toN-[(2S,3S,4R)-3,4-dihydroxy-1-{[(2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy-}octadecan-2-yl]-11-(3-methyloxetan-3-yl)undecanamide,except using THF a solvent. Purified by column chromatography on silicagel (PE/EtOAc 3:1) to giveN-((2S,3S,4R)-3,4-bis(benzyloxy)-1-(((2S,3R,4S,5S,6R)-3,4,5-tris(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-7-(3-(tetrahydro-2H-pyran-4-yl)propoxy)heptanamide(120 mg, 51%) as a solid. LC/MS: mass calcd. for C₈₁H₁₁₁NO₁₁: 1273.82,found: 1274.50 [M+H]⁺.

Step 3: Synthesis ofN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-7-(3-(tetrahydro-2H-pyran-4-yl)propoxy)heptanamide

A mixture ofN-((2S,3S,4R)-3,4-bis(benzyloxy)-1-(((2S,3R,4S,5S,6R)-3,4,5-tris(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-7-(3-(tetrahydro-2H-pyran-4-yl)propoxy)heptanamide(24 mg, 0.019 mmol), Pd(OH)₂ (40 mg), EtOH (3 mL) and DCM (3 mL) wasstirred under an atmosphere of H₂ (balloon) for 16 h. The mixture wasfiltered through a pad of Celite and the filtrate was concentrated underreduced pressure to giveN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-7-(3-(tetrahydro-2H-pyran-4-yl)propoxy)heptanamide(7.1 mg, 51%) as a solid. LC/MS: mass calcd. for C₃₉H₇₅NO₁₁: 733.53,found: 734.50 [M+H]⁺; ¹H NMR (400 MHz, CD₃OD) δ 4.23 (q, J=4.9 Hz, 1H),3.57-3.96 (m, 10H), 3.40-3.46 (m, 5H), 2.24 (t, J=7.8 Hz, 2H), 1.55-1.68(m, 9H), 1.21-1.41 (m, 37H), 0.88-0.96 (m, 3H).

Synthesis ofN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-6-(4-(tetrahydro-2H1-pyran-4-yl)butoxy)hexanamide

Step 1: Synthesis of 6-(4-(tetrahydro-2H-pyran-4-yl)butoxy)hexanoic Acid

Prepared in a manner similar to7-(3-(tetrahydro-2H-pyran-4-yl)propoxy)heptanoic acid and purified bycolumn chromatography on silica gel (PE/EtOAc 2:1) to give6-(4-(tetrahydro-2H-pyran-4-yl)butoxy)hexanoic acid (42 mg, 15%) as anoil. LC/MS: mass calcd. for C₁₅H₂₈O₄: 272, found 271 [M−H]⁻.

Step 2: Synthesis ofN-((2S,3S,4R)-3,4-bis(benzyloxy)-1-(((2S,3R,4S,5S,6R)-3,4,5-tris(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-6-(4-(tetrahydro-2H-pyran-4-yl)butoxy)hexanamide

Prepared in a manner similar toN-[(2S,3S,4R)-3,4-dihydroxy-1-{[(2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy}octadecan-2-yl]-11-(3-methyloxetan-3-yl)undecanamide,except using THF a solvent. Purified by column chromatography on silicagel (PE/EtOAc 3:1) to giveN-((2S,3S,4R)-3,4-bis(benzyloxy)-1-(((2S,3R,4S,5S,6R)-3,4,5-tris(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-6-(4-(tetrahydro-2H-pyran-4-yl)butoxy)hexanamide(20 mg, 10%) as a solid. LC/MS: mass calcd. for C₈₁H₁₁₁NO₁₁: 1273.82,found: 1274.50 [M+H]⁺.

Step 3: Synthesis ofN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-6-(4-(tetrahydro-2H-pyran-4-yl)butoxy)hexanamide

A mixture ofN-((2S,3S,4R)-3,4-bis(benzyloxy)-1-(((2S,3R,4S,5S,6R)-3,4,5-tris(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-6-(4-(tetrahydro-2H-pyran-4-yl)butoxy)hexanamide(26 mg, 0.02 mmol), Pd(OH)₂ (40 mg), EtOH (3 mL) and DCM (3 mL) wasstirred under an atmosphere of H₂ (balloon) for 16 h. The mixture wasfiltered through a pad of Celite and the filtrate was concentrated underreduced pressure to giveN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-6-(4-(tetrahydro-2H-pyran-4-yl)butoxy)hexanamide(1.3 mg, 8.5%) as an oil. LC/MS: mass calcd. for C₃₉H₇₅NO₁₁: 733.53,found: 734.45 [M+H]⁺; ¹H NMR (400 MHz, CD₃OD) δ 4.23 (q, J=5.0 Hz, 1H),3.98-3.77 (m, 10H), 3.40-3.47 (m, 5H), 2.26 (t, J=7.6 Hz, 2H), 1.52-1.68(m, 10H), 1.21-1.41 (m, 36H), (0.88-0.96 (m, 3H).

Synthesis ofN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-21-pyran-2-yl)oxy)octadecan-2-yl)-17-(3-fluorobicyclo[1.1.1]pentan-1-yl)heptadecanamide

Step 1: Synthesis of (15-carboxypentadecyl)triphenylphosphonium bromide

To a mixture of 16-bromohexadecanoic acid (2.0 g, 5.9 mmol) and CH₃CN(30 mL) was added Ph₃P (1.56 g, 5.9 mmol). The mixture was heated to 90°C., and stirred for 2 days, then filtered and washed with THF to give(15-carboxypentadecyl)triphenylphosphonium bromide (3.0 g, 84%) as asolid.

Step 2: Synthesis of17-(3-fluorobicyclo[1.1.1]pentan-1-yl)heptadec-16-enoic Acid

To a mixture of (9-carboxynonyl)triphenylphosphonium bromide (800 mg,1.3 mmol) in THF (15 mL) at 0° C. under an atmosphere of N₂ was added 2M NaHMDS (1.65 mL, 3.3 mmol). The mixture was warmed to rt and stirredfor 1 h, then 3-fluorobicyclo[1.1.1]pentane-1-carbaldehyde (152 mg, 1.3mmol) in THF (1 mL) was added. The mixture was stirred at rt overnight,then acidified pH˜1 with 1N HCl and extracted with EtOAc (3×30 mL). Thecombined organic layers were washed with brine (30 mL), dried overanhydrous Na₂SO₄, filtered, and the filtrate was concentrated underreduced pressure. The residue was purified by column chromatography onsilica gel (PE/EtOAc 2:1) to give17-(3-fluorobicyclo[1.1.1]pentan-1-yl)heptadec-16-enoic acid (140 mg,30%) as a solid. LC/MS: mass calcd. for C₂₂H₃₇FO₂: 352, found: 351[M−H]⁻.

Step 3: Synthesis of 17-(3-fluorobicyclo[1.1.1]pentan-1-yl)heptadecanoicAcid

A mixture of 17-(3-fluorobicyclo[1.1.1]pentan-1-yl)heptadec-16-enoicacid (140 mg, 0.39 mmol). PtO₂ (20 mg, 0.1 mmol) and EtOH (50 mL) wasstirred under an atmosphere of H₂ (balloon) for 1 h. The mixture wasfiltered through a pad of Celite and the filtrate was concentrated underreduced pressure to give17-(3-fluorobicyclo[1.1.1]pentan-1-yl)heptadecanoic acid (140 mg, 99%)as a solid. LC/MS: mass calcd. for C₂₂H₃₉FO₂: 354, found: 353 [M−H]⁻.

Step 4: Synthesis ofN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-17-(3-fluorobicyclo[1.1.1]pentan-1-yl)heptadecanamide

Prepared in a manner similar toN-[(2S,3S,4R)-3,4-dihydroxy-1-{[(2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy}octadecan-2-yl]-11-(3-methyloxetan-3-yl)undecanamide,purified by column chromatography on silica gel (DCM/MeOH 5:1) andpreparative-HPLC to giveN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-17-(3-fluorobicyclo[1.1.1]pentan-1-yl)heptadecanamide(18.8 mg, 15.7%) as a solid. LC/MS: mass calcd. for C₄₆H₈₆FNO₉: 815.63,found: 816.65 [M+H]⁺; ¹H NMR (400 M MHz, CD₃OD) δ 4.20 (dt, J=6.7, 4.4Hz, 1H), 3.73-3.93 (m, 5H), 3.53-3.76 (m, 5H), 2.24 (t, J=7.5 Hz, 2H),1.88 (d, J=2.6 Hz, 6H), 1.58-1.66 (m, 6H), 1.30-1.36 (m, 50H), 0.96-0.88(m, 3H); ¹⁹F NMR (376 MHz, CD₃PD) δ−146.5.

Synthesis ofN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-23-(3-fluorobicyclo[1.1.1]pentan-1-yl)tricosanamide

Step 1: Synthesis of (21-carboxyhenicosyl)triphenylphosphonium Bromide

To a mixture of 22-bromodocosanoic acid (1.4 g, 3.4 mmol) and CH₃CN (30mL) under an atmosphere of N₂ was added Ph₃P (0.91 g, 3.4 mmol). Themixture was heated to 90° C., and stirred for 2 days, then concentratedunder reduced pressure to give (21-carboxyhenicosyl)triphenylphosphoniumbromide (1.6 g, 68%) as a solid.

Step 2: Synthesis of23-(3-fluorobicyclo[1.1.1]pentan-1-yl)tricos-22-enoic Acid

To a mixture of (21-carboxyhenicosyl)triphenylphosphonium bromide (300mg, 0.4 mmol) in 1,4-dioxane (10 mL) under an atmosphere of N₂ at rt wasadded 4A molecular sieve (0.5 g), K₂CO₃ (245 mg, 1.7 mmol) and3-fluorobicyclo[1.1.1]pentane-1-carbaldehyde (75 mg, 0.7 mmol). Themixture was heated to 100° C. and stirred for 16 h, then diluted withH₂O, the mixture adjusted to pH 4˜5 with 2N HCl and extracted withEtOAc. The combined organic layers were concentrated under reducedpressure and the residue was purified by column chromatography on silicagel (PE/EtOAc 2:1) to give23-(3-fluorobicyclo[1.1.1]pentan-1-yl)tricos-22-enoic acid (50 mg, 26%)as a solid. LC/MS: mass calcd. for C₂₅H₄₉FO₂: 436, found: 435 [M−H]⁻.

Step 3: Synthesis of 23-(3-fluorobicyclo[1.1.1]pentan-1-yl)tricosanoicAcid

A mixture of 23-(3-fluorobicyclo[1.1.1]pentan-1-yl)tricos-22-enoic acid(50 mg, 0.11 mmol) and PtO₂ (5 mg) in EtOH (20 mL) was stirred at rtunder an atmosphere of H₂ (1 atm) for 1 h. The mixture was filteredthrough a pad of Celite and the filter cake washed with EtOH. Thefiltrate was concentrated under reduced pressure to give23-(3-fluorobicyclo[1.1.1]pentan-1-yl)tricosanoic acid (45 mg, 90%) as asolid. LC/MS: mass calcd. for C₂₈H₅₁FO₂: 438, found: 437 [M−H]⁻.

Step 4: Synthesis ofN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-23-(3-fluorobicyclo[1.1.1]pentan-1-yl)tricosanamide

Prepared in a manner similar toN-[(2S,3S,4R)-3,4-dihydroxy-1-{[(2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy}octadecan-2-yl]-11-(3-methyloxetan-3-yl)undecanamide,except DMF/THF was used as solvent. Purified by column chromatography onsilica gel (DCM/MeOH) and washed with CH₃CN to giveN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-23-(3-fluorobicyclo[1.1.1]pentan-1-yl)tricosanamide(5.2 mg, 5.2%) as a solid. LC/MS: mass calcd. for C₅₂H₉₈FNO₉: 899.72,found: 922.65 [M+H]⁺; ¹H NMR (300 MHz, CD₃OD) δ 4.13-4.25 (m, 1H),3.56-3.93 (m, 10H), 2.24 (t, J=7.4 Hz, 2H), 1.88 (d, J=2.6 Hz, 6H),1.55-1.68 (m, 5H), 1.26-1.42 (m, 63H), 0.92 (t, J=6.7 Hz, 3H); ¹⁹F NMR(282 MHz, CD₃OD) δ−146.5.

Synthesis ofN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-21-(3-fluorobicyclo[1.1.1]pentan-1-yl)heneicosanamine

Step 1: Synthesis of21-(3-fluorobicyclo[1.1.1]pentan-1-yl)henicos-20-enoic Acid

Prepared in a manner similar to23-(3-fluorobicyclo[1.1.1]pentan-1-yl)tricos-22-enoic acid and purifiedby column chromatography on silica gel (PE/EtOAc 1:1) to give21-(3-fluorobicyclo[1.1.1]pentan-1-yl)henicos-20-enoic acid (65 mg, 35%)as a solid. LC/MS: mass calcd. for C₂₆H₄₅FO₂: 408, found: 407 [M−H]⁻.

Step 2: Synthesis of 21-(3-fluorobicyclo[1.1.1]pentan-1-yl)heneicosanoicAcid

Prepared in a similar manner to23-(3-fluorobicyclo[1.1.1]pentan-1-yl)tricosanoic acid to give21-(3-fluorobicyclo[1.1.1]pentan-1-yl)heneicosanoic acid (60 mg, 92%) asa solid. LC/MS: mass calcd. for C₂₆H₄₇FO₂: 41, found: 409 [M−H]⁻.

Step 3: Synthesis ofN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-21-(3-fluorobicyclo[1.1.1]pentan-1-yl)heneicosanamine

Prepared in a manner similar toN-[(2S,3S,4R)-3,4-dihydroxy-1-{[(2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy}octadecan-2-yl]-11-(3-methyloxetan-3-yl)undecanamide,except DMF/THF was used as solvent. Purified by column chromatography onsilica gel (DCM/MeOH 9:1) and washed with CH₃CN to giveN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-21-(3-fluorobicyclo[1.1.1]pentan-1-yl)heneicosanamine(4.7 mg, 3.9%) as a solid. LC/MS: mass calcd. for C₅₀H₉₄FNO₉: 871.69,found: 872.70 [M+H]⁺ and 894.65 [M+Na]⁺; ¹H NMR (300 MHz, CD₃OD) δ4.16-4.23 (m, 1H), 3.56-3.89 (m, 10H), 2.24 (t, J=7.5 Hz, 2H), 1.88 (d,J=2.6 Hz, 6H), 1.52-1.69 (m, 6H), 1.23-1.39 (m, 58H), 0.92 (t, J=6.4 Hz,3H); ¹⁹F NMR (282 MHz, CD₃OD) δ−146.6.

Synthesis ofN-[(2S,3S,4R)-3,4-dihydroxy-1-{[(2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy}octadecan-2-yl]-22-{3-fluorobicyclo[1.1.1]pentan-1-yl}docosanamide

Step 1: Synthesis of 21-(bromotriphenyl-lambda5-phosphanyl)heneicosanoicAcid

To a mixture of 21-bromohenicosanoic acid (1.16 g, 2.86 mmol) and CH₃CN(20 mL) was added Ph₃P (0.75 g, 2.86 mmol). The mixture was heated to90° C., and stirred for 2 days, then concentrated under reducedpressure. The residue was purified by C18 reverse-phase HPLC (H₂O, 5%HCl)/CH₃OH 5%-100%) to give21-(bromotriphenyl-lambda5-phosphanyl)heneicosanoic acid (1.27 g, 66%)as a solid.

Step 2: Synthesis of(E)-22-(3-fluorobicyclo[1.1.1]pentan-1-yl)docos-21-enoic Acid

Prepared in a manner similar to23-(3-fluorobicyclo[1.1.1]pentan-1-yl)tricos-22-enoic acid and purifiedby column chromatography on silica gel (PE/EtOAc 1:1) to give(E)-22-(3-fluorobicyclo[1.1.1]pentan-1-yl)docos-21-enoic acid (44 mg,35%) as a solid. LC/MS: mass calcd. for C₂₇H₄₇FO₂: 422.36, found: 421.15[M−H]⁻.

Step 3: Synthesis of 22-(3-fluorobicyclo[1.1.1]pentan-1-yl)docosanoicAcid

Prepared in a similar manner to23-(3-fluorobicyclo[1.1.1]pentan-1-yl)tricosanoic acid to give22-(3-fluorobicyclo[1.1.1]pentan-1-yl)docosanoic acid (40 mg, 99%) as asolid. LC/MS: mass calcd. for C₂₇H₄₉FO₂: 424.37, found: 423.15 [M−H]⁻.

Step 4: Synthesis ofN-[(2S,3S,4R)-3,4-dihydroxy-1-{[(2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy}octadecan-2-yl]-22-{3-fluorobicyclo[1.1.1]pentan-1-yl}docosanamide

Prepared in a manner similar toN-[(2S,3S,4R)-3,4-dihydroxy-1-{[(2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy}octadecan-2-yl]-11-(3-methyloxetan-3-yl)undecanamide,except DMF/THF was used as solvent. Purified by column chromatography onsilica gel (DCM/MeOH 9:1) and washed with CH₃CN to giveN-[(2S,3S,4R)-3,4-dihydroxy-1-{[(2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy}octadecan-2-yl]-22-{3-fluorobicyclo[1.1.1]pentan-1-yl}docosanamide(22.5 mg, 18%) as a solid. LC/MS: mass calcd. for C₅₁H₉₆FNO₉: 885.71,found: 886.75 [M+H]⁺; ¹H NMR (400 MHz, CD₃OD) δ 4.85-4.87 (m, 1H), 4.20(d, J=6.6 Hz, 1H), 3.89-3.90 (m, 3H), 3.70-3.85 (m, 5H), 3.63-3.67 (m,1H), 3.55-3.60 (m, 1H), 2.24 (t, J=7.5 Hz, 2H), 1.88 (d, J=2.6 Hz, 6H),1.56-1.66 (m, 4H), 1.30-1.46 (m, 62H), 0.92 (t, J=6.8 Hz, 3H); ¹⁹F NMR(376 MHz, CD₃OD) δ−146.6.

Synthesis ofN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-24-(3-fluorobicyclo[1.1.1]pentan-1-yl)tetracosanamide

Step 1: Synthesis of 8-(3-fluorobicyclo[1.1.1]pentan-1-yl)oct-7-enoicAcid

Prepared in a manner similar to17-(3-fluorobicyclo[1.1.1]pentan-1-yl)heptadec-16-enoic acid andpurified by column chromatography on silica gel (PE/EtOAc 1:1) to give8-(3-fluorobicyclo[1.1.1]pentan-1-yl)oct-7-enoic acid (500 mg, 50%) as asolid. LC/MS: mass calcd. for C₁₃H₁₉FO₂: 226, found: 225 [M−H]⁻.

Step 2: Synthesis of 8-{3-fluorobicyclo[1.1.1]pentan-1-yl}octanoic Acid

A mixture of 8-(3-fluorobicyclo[1.1.1]pentan-1-yl)oct-7-enoic acid (500mg, 2.21 mmol) and PtO₂ (30 mg) in EtOH (100 mL) was stirred under anatmosphere of H₂ (balloon) for 1 h. The mixture was filtered through apad of Celite and the filtrate was concentrated under reduced pressureto give 8-{3-fluorobicyclo[1.1.1]pentan-1-yl}octanoic acid (500 mg, 99%)as a solid. LC/MS: mass calcd. for C₁₃H₂₁FO₂: 228, found: 227 [M−H]⁻.

Step 3: Synthesis of 8-{3-fluorobicyclo[1.1.1]pentan-1-yl}octan-1-ol

To a mixture of 8-{3-fluorobicyclo[1.1.1]pentan-1-yl}octanoic acid (500mg, 2.19 mmol) and THF (10 mL) at 0° C. under an atmosphere of N₂ wasadded LiAlH₄ (332 mg, 8.76 mmol). The mixture was stirred at 0° C. for 3h, then diluted with aqueous Na₂SO₄ and filtered. The filtrate waswashed with EtOAc, and the organic layer was dried over Na₂SO₄, filteredand the filtrate was concentrated under reduced pressure. The residuewas purified by column chromatography on silica gel (PE/EtOAc 1:1) togive 8-{3-fluorobicyclo[1.1.1]pentan-1-yl}octan-1-ol (320 mg, 68%) as asolid.

Step 4: Synthesis of 8-(3-fluorobicyclo[1.1.1]pentan-1-yl)octanal

To a mixture of oxalyl chloride (284 mg, 2.24 mmol) and DCM (10 mL) at−78° C. under an atmosphere of N₂ was added DMSO (349 mg, 4.47 mmol)dropwise. The mixture was stirred at −78° C. for 15 min. then8-{3-fluorobicyclo[1.1.1]pentan-1-yl}octan-1-ol (320 mg, 1.49 mmol) inDCM (2 mL) was added dropwise. The mixture was stirred at −78° C. for 50min, then Et₃N (3 mL) was added. Stirring was continued at −78° C. for 5min, then the mixture was warmed to rt, diluted with H₂O (10 mL) andextracted with DCM (3×20 mL). The combined organic layers were washedwith H₂O (20 mL), brine (20 mL), dried over Na₂SO₄, and filtered. Thefiltrate was concentrated under reduced pressure to give8-(3-fluorobicyclo[1.1.1]pentan-1-yl)octanal (300 mg, 95%) as an oil,which was used without further purification R_(f)=0.4 (PE/EtOAc=1:2).

Step 5: Synthesis of24-(3-fluorobicyclo[1.1.1]pentan-1-yl)tetracos-16-enoic Acid

To a mixture of (15-carboxy pentadecyl)triphenylphosphonium bromide (844mg, 1.41 mmol) and THF (20 mL) at 0° C. under an atmosphere of N₂ wasadded NaHMDS (1.5 mL, 9.6 mmol). The mixture was warmed to rt andstirred for 1 h, then 8-{3-fluorobicyclo[1.1.1]pentan-1-yl}octanal (300mg, 1.41 mmol) was added and the mixture was stirred at rt overnight.The mixture was acidified to pH˜5 with 1N HCl and extracted with EtOAc(3×30 mL). The combined organic layers were washed with brine (50 mL),dried over anhydrous Na₂SO₄, filtered and the filtrate was concentratedunder reduced pressure. The residue was purified by columnchromatography on silica gel (PE/EtOAc 1:1) to give24-(3-fluorobicyclo[1.1.1]pentan-1-yl)tetracos-16-enoic acid (400 mg,63%) as a solid. LC/MS-mass calcd. for C29H51FO₂: 450, found: 449[M−H]⁻.

Step 6: Synthesis of 24-(3-fluorobicyclo[1.1.1]pentan-1-yl)tetracosanoicAcid

Prepared in a similar manner to23-(3-fluorobicyclo[1.1.1]pentan-1-yl)tricosanoic acid to give24-(3-fluorobicyclo[1.1.1]pentan-1-yl)tetracosanoic acid (180 mg, 90%)as a solid. LC/MS: mass calcd. for C₂₉H₅₃FO₂: 452, found: 451 [M−H]⁻.

Step 7: Synthesis ofN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-24-(3-fluorobicyclo[1.1.1]pentan-1-yl)tetracosanamide

Prepared in a manner similar toN-[(2S,3S,4R)-3,4-dihydroxy-1-{[(2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy}octadecan-2-yl]-11-(3-methyloxetan-3-yl)undecanamide,except DMF/THF was used as solvent. Purified by column chromatography onsilica gel (DCM/MeOH 9:1) and washed with CH₃CN to giveN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-24-(3-fluorobicyclo[1.1.1]pentan-1-yl)tetracosanamide(12.0 mg, 9.4%). LC/MS: mass calcd. for C₅₃H₁₀₀FNO₉: 913.74, found:936.80 [M+Na]⁺; ¹H NMR (400 MHz, CD₃OD) δ 4.20 (d, J=6.4 Hz, 1H),3.53-3.93 (m, 10H), 2.09-2.36 (m, 2H), 1.88 (d, J=2.7 Hz, 6H), 1.56-1.67(m, 6H), 0.92 (t, J=6.6 Hz, 4H); ¹⁹F NMR (376 MHz, CD₃OD) δ−146.5.

Synthesis ofN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-25-(3-fluorobicyclo[1.1.1]pentan-1-yl)pentacosanamide

Step 1: Synthesis of 9-(3-fluorobicyclo[1.1.1]pentan-1-yl)non-8-enoicAcid

Prepared in a manner similar to1743-fluorobicyclo[1.1.1]pentan-1-yl)heptadec-16-enoic acid and purifiedby column chromatography on silica gel (PE/EtOAc 1:1) to give9-(3-fluorobicyclo[1.1.1]pentan-1-yl)non-8-enoic acid (650 mg, 62%) as asolid. LC/MS: mass calcd. for C₁₄H₂₁FO₂: 240, found: 239 [M−H]⁻.

Step 2: Synthesis of 9-{3-fluorobicyclo[1.1.1]pentan-1-yl}nonanoic Acid

Prepared in a manner similar to8-{3-fluorobicyclo[1.1.1]pentan-1-yl}octanoic acid to give9-{3-fluorobicyclo[1.1.1]pentan-1-yl}nonanoic acid (650 mg, 99%) as asolid. LC/MS: mass calcd. for C₁₄H₂₃FO₂: 242, found: 240 [M−H]⁻.

Step 3: Synthesis of 9-{3-fluorobicyclo[1.1.1]pentan-1-yl}nonan-1-ol

Prepared in a similar manner to8-{3-fluorobicyclo[1.1.1]pentan-1-yl}octan-1-ol and purified by columnchromatography on silica gel (PE/EtOAc 1:1) to give9-{3-fluorobicyclo[1.1.1]pentan-1-yl}nonan-1-ol (400 mg, 65%) as asolid.

Step 4: Synthesis of 9-{3-fluorobicyclo[1.1.1]pentan-1-yl}nonanal

Prepared in a similar manner to8-(3-fluorobicyclo[1.1.1]pentan-1-yl)octanal to give9-{3-fluorobicyclo[1.1.1]pentan-1-yl}nonanal (280 mg, 94%) R_(f)=0.4.PE/EA=1:3.

Step 5: Synthesis of25-(3-fluorobicyclo[1.1.1]pentan-1-yl)pentacos-16-enoic Acid

Prepared in a similar manner to24-(3-fluorobicyclo[1.1.1]pentan-1-yl)tetracos-16-enoic acid to give andpurified by column chromatography on silica gel (PE/EtOAc 1:1) to give25-(3-fluorobicyclo[1.1.1]pentan-1-yl)pentacos-16-enoic acid (200 mg,35%) as a solid. LC/MS: mass calcd. for C₃₀H₅₃FO₂: 464, found: 463[M−H]⁻.

Step 6: Synthesis of 25-{3-fluorobicyclo[1.1.1]pentan-1-yl}pentacosanoicAcid

A mixture of 25-{3-fluorobicyclo[1.1.1]pentan-1-yl}pentacos-16-enoicacid (200 mg, 0.43 mmol) and PtO₂ (30 mg) in EtOH (100 mL) was stirredunder an atmosphere of H₂ (balloon) for 1 h. The reaction mixture wasfiltered through a pad of celite and concentrated under reduced pressureto afford 25-{3-fluorobicyclo[1.1.1]pentan-1-yl}pentacosanoic acid (200mg, 99.6%) as a white solid. LC/MS: mass calcd. for C₃₀H₅₅FO₂: 466,found: 465 [M−H]⁻.

Step 7: Synthesis ofN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-25-(3-fluorobicyclo[1.1.1]pentan-1-yl)pentacosanamide

Prepared in a manner similar toN-[(2S,3S,4R)-3,4-dihydroxy-1-{[(2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy}octadecan-2-yl]-11-(3-methyloxetan-3-yl)undecanamide,except DMF/THF was used as solvent. Purified by column chromatography onsilica gel (DCM/MeOH 5:1) and preparative-HPLC to giveN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-25-(3-fluorobicyclo[1.1.1]pentan-1-yl)pentacosanamide(8.8 mg, 7.7%). LC/MS: mass calcd. for C₅₄H₁₀₂FNO₉: 927.75, found:950.85 [M+Na]⁺; ¹H NMR (300 MHz, CD₃OD) δ 4.20 (d, J=5.5 Hz, 1H),3.54-3.93 (m, 10H), 2.24 (t, J=7.4 Hz, 2H), 1.88 (d, J=2.7 Hz, 6H),1.54-1.69 (m, 6H), 1.25-1.42 (m, 66H), 0.91 (t, J=8.6, 7.4 Hz, 3H); ¹⁹FNMR (282 MHz, CD₃OD) δ−146.6.

Synthesis ofN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-11-(3-tridecylbicyclo[1.1.1]pentan-1-yl)undecanamide

Step 1: Synthesis oftert-butyldiphenyl((3-(tridec-1-en-1-yl)bicyclo[1.1.1]pentan-1-yl)methoxy)silane

To a mixture of dodecyltriphenylphosphonium bromide (2.1 g, 4.1 mmol)and THF (20 mL) at −78° C. under an atmosphere of N₂ was added n-BuLi(1.8 mL, 4.5 mmol). The mixture was stirred at −78° C. for 20 min,warmed to 0° C., and stirred 20 min, then a solution of3-(((tert-butyldiphenylsilyl)oxy)methyl)bicyclo[1.1.1]pentane-1-carbaldehyde[CAS No: 1678528-05-6] (1.5 g, 4.1 mmol) in THF (5 mL). The mixture waswarmed to rt and stirred overnight, then diluted with H₂O (30 mL) andextracted with EtOAc (3×50 mL). The combined organic layers were washedwith brine (50 mL), dried over anhydrous Na₂SO₄, filtered and thefiltrate was concentrated under reduced pressure. The residue waspurified by column chromatography on silica gel (PE/EtOAc 4:1) to givetert-butyldiphenyl((3-(tridec-1-en-1-yl)bicyclo[1.1.1]pentan-1-yl)methoxy)silane(1.3 g, 61%) as a solid.

Step 2: Synthesis oftert-butyldiphenyl((3-tridecylbicyclo[1.1.1]pentan-1-yl)methoxy)silane

A mixture oftert-butyldiphenyl((3-(tridec-1-en-1-yl)bicyclo[1.1.1]pentan-1-yl)methoxy)silane(1.3 g, 2.5 mmol), PtO₂ (100 mg) and EtOH (100 mL) was stirred at rtunder an atmosphere of H₂ (1 atm) for 1 h. The mixture was filteredthrough a pad of Celite and the filter cake was washed with EtOH. Thefiltrate was concentrated under reduced pressure to givetert-butyldiphenyl((3-tridecylbicyclo[1.1.1]pentan-1-yl)methoxy)silane(1.1 g, 84%) as a solid.

Step 3: Synthesis of (3-tridecylbicyclo[1.1.1]pentan-1-yl)methanol

To a mixture oftert-butyldiphenyl((3-tridecylbicyclo[1.1.1]pentan-1-yl)methoxy)silane(1.1 g, 2.1 mmol) and THF (30 mL) at rt was added TBAF (1.66 g, 6.4mmol). The mixture was stirred at rt for 3 h, then extracted with EtOAc(3×20 mL), The combined organic layers were washed with brine (50 mL),dried over anhydrous Na₂SO₄, filtered and the filtrate was concentratedunder reduced pressure. The residue was purified by columnchromatography on silica gel (PE/EtOAc 1:1) to give(3-tridecylbicyclo[1.1.1]pentan-1-yl)methanol (500 mg, 84%) as a solid.

Step 4: Synthesis of 3-tridecylbicyclo[1.1.1]pentane-1-carbaldehyde

DMSO (267 mg, 3.4 mmol) was added dropwise to a mixture of oxalylchloride (217 mg, 1.7 mmol) at −78° C. under an atmosphere of N₂. Themixture was stirred for 15 mm-78° C., then3-tridecylbicyclo[1.1.1]pentan-1-yl)methanol (320 mg, 1.2 mmol) in DCM(2 mL) was added dropwise and the mixture was stirred at −78° C. for 50min. Et₃N (3 mL) was added at −78° C., and stirring was continued for anadditional 5 min, then the mixture was warmed to rt and diluted with H₂O(10 mL) and extracted with DCM (3×20 mL). The combined organic layerswere washed with H₂O (20 mL), brine (20 mL), dried over Na₂SO₄, filteredand the filtrate concentrated under reduced pressure to give3-tridecylbicyclo[1.1.1]pentane-1-carbaldehyde (300 mg, 94%) as an oil.The product was used without further purification.

Step 5: Synthesis of11-(3-tridecylbicyclo[1.1.1]pentan-1-yl)undec-10-enoic Acid

To a mixture of 10-(bromotriphenyl-lambda5-phosphanyl)decanoic acid (553mg, 1.0 mmol) and THF (20 mL) at 0° C. under an atmosphere of N₂ wasadded NaHMDS (1.2 mL, 2.4 mmol). The mixture was warmed to rt andstirred for 1 h, then a mixture of3-tridecylbicyclo[1.1.1]pentane-1-carbaldehyde (300 mg, 1.0 mmol) in THF(2 mL) was added and the mixture was stirred at rt overnight. Themixture was acidified to pH˜5 with 1N HCl and extracted with EtOAc (3×30mL), The combined organic layers were washed with brine (30 mL), driedover anhydrous Na₂SO₄, filtered and the filtrate was concentrated underreduced pressure. The residue was purified by column chromatography onsilica gel eluted (PE/EtOAc 1:1) to give11-(3-tridecylbicyclo[1.1.1]pentan-1-yl)undec-10-enoic acid (160 mg,34%) as a solid.

Step 6: Synthesis of 11-(3-tridecylbicyclo[1.1.1]pentan-1-yl)undecanoicAcid

A mixture of 11-{3-tridecylbicyclo[1.1.1]pentan-1-yl}undec-10-enoic acid(160 mg, 0.43 mmol), PtO₂ (25 mg) and EtOH (50 mL) was stirred at rtunder an atmosphere of H₂ (1 atm) for 1 h. The mixture was filteredthrough a pad of Celite and the filter cake was washed with EtOH. Thefiltrate was concentrated under reduced pressure to give11-(3-tridecylbicyclo[1.1.1]pentan-1-yl)undecanoic acid (150 mg, 93%) asa solid.

Step 7: Synthesis ofN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H1-pyran-2-yl)oxy)octadecan-2-yl)-11-(3-tridecylbicyclo[1.1.1]pentan-1-yl)undecanamide

To a mixture of 11-{3-tridecylbicyclo[1.1.1]pentan-1-yl}undecanoic acid(45 mg, 0.10 mmol) and(2S,3R,4S,5R,6R)-2-(((2S,3S,4R)-2-amino-3,4-dihydroxyoctadecyl)oxy)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol(49 mg, 0.10 mmol) in DMF (3 mL) and THF (3 mL) at rt under anatmosphere of N₂ was added HBTU (118 mg, 0.31 mmol), Et₃N (0.1 mL) andNMM (0.1 mL). The mixture was stirred at rt for 16 h, then diluted withH₂O (10 mL) and extracted with EtOAc (30 mL×3). The combined organiclayers were washed with brine (30 mL×2), dried over Na₂SO₄, filtered andthe filtrate was concentrated under reduced pressure. The residue waspurified by column chromatography on silica gel (DCM/MeOH) and washedwith CH₃CN to giveN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-11-(3-tridecylbicyclo[1.1.1]pentan-1-yl)undecanamide(15.3 mg, 15.7%). LC/MS: mass calcd. for C₅₃H₁₀₁NO₉: 895.75, found:918.75 [M+Na]⁺; ¹H NMR (400 MHz, CD₃OD) δ 4.17 (d, J=6.8, 4.2 Hz, 1H),3.50-3.90 (m, 10H), 2.22 (t, J=7.5 Hz, 2H), 1.64-1.57 (m, 6H), 1.43 (s,6H), 1.29-1.41 (m, 62H), 0.88-0.91 (m, 6H).

Synthesis ofN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-17-(oxetan-3-yl)heptadecanamide

Step 1: Synthesis of(E)-22-(3-fluorobicyclo[1.1.1]pentan-1-yl)docos-21-enoic Acid

Prepared in a manner similar to17-(3-fluorobicyclo[1.1.1]pentan-1-yl)heptadec-16-enoic and purified bycolumn chromatography on silica gel (PE/EtOAc 1:1) to give17-(oxetan-3-yl)heptadec-16-enoic acid (60 mg, 37%) as a solid. LC/MS:mass calcd. for C₂₀H₃₆O₃: 324, found: 323 [M−H]⁻.

Step 2: Synthesis of 22-(3-fluorobicyclo[1.1.1]pentan-1-yl)docosanoicAcid

Prepared in a similar manner to17-(3-fluorobicyclo[1.1.1]pentan-1-yl)heptadecanoic acid to give17-(oxetan-3-yl)heptadecanoic acid (60 mg, 99%) as a solid. LC/MS: masscalcd. for C₂₀H₃₈O₃: 326, found: 325 [M−H]⁻.

Step 3: Synthesis ofN-[(2S,3S,4R)-3,4-dihydroxy-1-{[(2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy}octadecan-2-yl]-22-{3-fluorobicyclo[1.1.1]pentan-1-yl}docosanamide

Prepared in a manner similar toN-[(2S,3S,4R)-3,4-dihydroxy-1-{[(2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy}octadecan-2-yl]-11-(3-methyloxetan-3-yl)undecanamide,except DMF/THF was used as solvent. Purified by column chromatography onsilica gel (DCM/MeOH) and washed with CH₃CN to giveN-((2S,3S,4R)-3,4-dihydroxy-1-((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-17-(oxetan-3-yl)heptadecanamide(17.5 mg, 15%) as a solid. LC/MS: mass calcd. for C₄₄H₈₅NO₁₀: 787.62,found: 810.55 [M+Na]⁺; ¹H NMR (400 MHz, CD₃OD) δ 4.81 (dd, J=7.8, 5.8Hz, 2H), 4.39 (t, J=6.1 Hz, 2H), 4.20 (q, J=4.6 Hz, 1H), 3.93-3.81 (m,3H), 3.85-3.60 (m, 6H), 3.57 (t, J=7.4 Hz, 1H), 3.07-2.95 (m, 1H), 2.24(t, J=7.5 Hz, 2H), 1.58-1.74 (m, 6H), 1.26-1.40 (m, 50H), 0.92 (t, J=6.7Hz, 3H).

Synthesis ofN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-21-(oxetan-3-yl)heneicosanamine

Step 1: Synthesis of 21-(oxetan-3-yl)henicos-20-enoic Acid

Prepared in a manner similar to23-(3-fluorobicyclo[1.1.1]pentan-1-yl)tricos-22-enoic acid purified bycolumn chromatography on silica gel (PE/EtOAc 2:1) to give21-(oxetan-3-yl)henicos-20-enoic acid (50 mg, 29%) as a solid. LC/MS:mass calcd. for C₂₄H₄₄O₃: 380.33, found: 379 [M−H]⁻.

Step 2: Synthesis of 21-(oxetan-3-yl)heneicosanoic Acid

Prepared in a similar manner to23-(3-fluorobicyclo[1.1.1]pentan-1-yl)tricosanoic acid to give21-(oxetan-3-yl)heneicosanoic acid (50 mg, 90%) as a solid LC/MS: masscalcd. for C₂₄H₄₆O₃: 382.34, found: 381 [M−H]⁻.

Step 3: Synthesis ofN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-21-(oxetan-3-yl)heneicosanamine

Prepared in a manner similar toN-[(2S,3S,4R)-3,4-dihydroxy-1-{[(2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy}octadecan-2-yl]-11-(3-methyloxetan-3-yl)undecanamide,except DMF/THF was used as solvent. Purified by column chromatography onsilica gel (DCM/MeOH) and washed with CH₃CN to giveN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-21-(oxetan-3-yl)heneicosanamine(5 mg, 4.4%) as a solid. LC/MS: mass calcd. for C₄₈H₉₃NO₁₀: 843.68,found: 866.65 [M+Na]⁺; ¹H NMR (300 MHz, CD₃OD) δ 4.81 (dd, J=7.8, 5.8Hz, 2H), 4.39 (t, J=6.1 Hz, 2H), 4.20 (d, J=6.0 Hz, 1H), 3.60-3.93 (m,9H), 2.24 (t, J=7.5 Hz, 2H), 1.57-1.79 (m, 6H), 1.13-1.38 (m, 60H),0.87-0.93 (m, 3H).

Synthesis ofN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-23-(oxetan-3-yl)tricosanamide

Step 1: Synthesis of 23-(oxetan-3-yl)tricos-22-enoic Acid

Prepared in a manner similar to23-(3-fluorobicyclo[1.1.1]pentan-1-yl)tricos-22-enoic acid purified bycolumn chromatography on silica gel (PE/EtOAc 2:1) to give23-(oxetan-3-yl)tricos-22-enoic acid (60 mg, 33%) as a solid. LC/MS:mass calcd. for C₂₆H₄₈O₃: 408.36: 408, found: 407 [M−H]⁻.

Step 2: Synthesis of 23-(oxetan-3-yl)tricosanoic Acid

Prepared in a similar manner to23-(3-fluorobicyclo[1.1.1]pentan-1-yl)tricosanoic acid to give23-(oxetan-3-yl)tricosanoic acid (55 mg, 91%) as a solid. LC/MS: masscalcd. for C₂₆H₅₀O₃: 410, found: 409 [M−H]⁻.

Step 3: Synthesis ofN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-23-(oxetan-3-yl)tricosanamide

Prepared in a manner similar toN-[(2S,3S,4R)-3,4-dihydroxy-1-{[(2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy}octadecan-2-yl]-11-(3-methyloxetan-3-yl)undecanamide,except DMF/THF was used as solvent. Purified by column chromatography onsilica gel (DCM/MeOH) and washed with CH₃CN to giveN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-23-(oxetan-3-yl)tricosanamide(5.2 mg, 4.3%) as a solid. LC/MS: mass calcd. for C₅₀H₉₇NO₁₀: 871.71,found: 894.60 [M+Na]⁺; ¹H NMR (400 MHz, CD₃OD) δ 4.81 (dd, J=7.8, 5.8Hz, 2H), 4.39 (t, J=6.1 Hz, 2H), 4.20 (d, J=6.8 Hz, 1H), 3.56-3.87 (m,10H), 2.24 (t, J=7.5 Hz, 2H), 1.54-1.71 (m, 6H), 1.24-1.38 (m, 64H),0.85-0.96 (m, 3H).

Synthesis ofN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-22-(oxetan-3-yl)docosanamide

Step 1: Synthesis of 22-(oxetan-3-ylidene)docosanoic Acid

Prepared in a manner similar to23-(3-fluorobicyclo[1.1.1]pentan-1-yl)tricos-22-enoic acid purified bycolumn chromatography on silica gel (PE/EtOAc 2:1) to give22-(oxetan-3-ylidene)docosanoic acid (60 mg, 35%) as a solid. LC/MS:mass calcd. for C₂₅H₄₆O₃: 394, found: 393 [M−H]⁻.

Step 2: Synthesis of 22-(oxetan-3-yl)docosanoic Acid

Prepared in a similar manner to23-(3-fluorobicyclo[1.1.1]pentan-1-yl)tricosanoic acid to give22-(oxetan-3-yl)docosanoic acid (55 mg, 91%) as a solid. LC/MS: masscalcd. for C₂₅H₄₈O₃: 396.36, found: 395 [M−H]⁻.

Step 3: Synthesis ofN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2l-pyran-2-yl)oxy)octadecan-2-yl)-22-(oxetan-3-yl)docosanamide

Prepared in a manner similar toN-[(2S,3S,4R)-3,4-dihydroxy-1-{[(2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy}octadecan-2-yl]-11-(3-methyloxetan-3-yl)undecanamide,except DMF/MIF was used as solvent. Purified by column chromatography onsilica gel (DCM/MeOH) and washed with CH₃CN to giveN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-22-(oxetan-3-yl)docosanamide(6.5 mg, 5.2%) as a solid. LC/MS: mass calcd. for C₄₉H₉₅NO₁₀: 857.70,found: 880.65 [M+Na]⁺; ¹H NMR (400 MHz, CD₃OD) δ 4.80-4.84 (m, 2H), 4.81(dd, J=7.8, 5.9 Hz, 2H), 4.16-4.22 (m, 1H), 3.54-3.96 (m, 10H),4.14-4.28 (m, 1H), 1.54-1.73 (m, 6H), 1.28-1.41 (m, 61H), 0.86-0.99 (m,3H).

Synthesis ofN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-21-pyran-2-yl)oxy)octadecan-2-yl)-24-(oxetan-3-yl)tetracosanamide

Step 1: Synthesis of (23-carboxytricosyl)triphenylphosphonium bromide

To a mixture of 24-bromotetracosanoic acid (0.6 g, 1.3 mmol) in CH₃CN(10 mL) under an atmosphere of N₂ was added triphenylphosphine (0.34 g,1.3 mmol, 1.0 equiv). The mixture was heated to 90° C., and stirred for2 days, then concentrated under reduced pressure and the residue waspurified by preparative-HPLC (H₂O 5% HCl/CH₃OH 5%-100%) to give(23-carboxytricosyl)triphenylphosphonium bromide (0.6 g, 70%) as asolid.

Step 2: Synthesis of 24-(oxetan-3-ylidene)tetracosanoic Acid

Prepared in a manner similar to23-(3-fluorobicyclo[1.1.1]pentan-1-yl)tricos-22-enoic acid purified bycolumn chromatography on silica gel (PE/EtOAc 2:1) to give24-(oxetan-3-ylidene)tetracosanoic acid (50 mg, 42%) as a solid. LC/MS:mass calcd. for C₂₇H₅₀O₃: 422, found: 421 [M−H]⁻.

Step 3: Synthesis of 24-(oxetan-3-yl)tetracosanoic Acid

Prepared in a similar manner to23-(3-fluorobicyclo[1.1.1]pentan-1-yl)tricosanoic acid to give24-(oxetan-3-yl)tetracosanoic acid (50 mg, 100%) as a solid. LC/MS: masscalcd. for C₂₇H₅₂O₃: 424, found: 423 [M−H]⁻.

Step 4: Synthesis ofN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-24-(oxetan-3-yl)tetracosanamide

Prepared in a manner similar toN-[(2S,3S,4R)-3,4-dihydroxy-1-{[(2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy}octadecan-2-yl]-11-(3-methyloxetan-3-yl)undecanamide,except DMF/THF was used as solvent. Purified by column chromatography onsilica gel (DCM/MeOH) and washed with CH₃CN to giveN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-24-(oxetan-3-yl)tetracosanamide(7 mg, 6.7%) as a solid. LC/MS: mass calcd. for C₅₁H₉₉NO₁₀: 885.73,found: 908.70 [M+Na]⁺; ¹H NMR (400 MHz, CD₃OD) δ 4.79-4.83 (m, 2H),4.37-4.42 (m, 2H), 4.17-4.21 (m, 1H), 3.56-3.91 (m, 10H), 2.21-2.28 (m,2H), 1.54-1.74 (m, 6H), 1.30-1.38 (m, 65H), 0.90-0.93 (m, 3H).

Synthesis ofN-[(2S,3S,4R)-3,4-dihydroxy-1-{[(2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy}octadecan-2-yl]-25-(oxetan-3-yl)pentacosanamide

Step 1: Synthesis of 9-(oxetan-3-yl)non-8-enoic Acid

Prepared in a manner similar to17-(3-fluorobicyclo[1.1.1]pentan-1-yl)heptadec-16-enoic purified bycolumn chromatography on silica gel (PE/EtOAc 1:1) to give9-(oxetan-3-yl)non-8-enoic acid (1.4 g, 58%) as a solid. LC/MS: masscalcd. for C₁₂H₂₀O₃: 212, found: 211 [M−H]⁺.

Step 2: Synthesis of 9-(oxetan-3-yl)nonanoic Acid

Prepared in a similar manner to17-(3-fluorobicyclo[1.1.1]pentan-1-yl)heptadecanoic acid to give9-(oxetan-3-yl)nonanoic acid (1.2 g, 100%) as a solid. LC/MS: masscalcd. for C₁₂H₂₂O₃: 214, found: 213 [M+H]⁺.

Step 3: Synthesis of 9-(oxetan-3-yl)nonan-1-ol

Prepared in a manner similar to8-{3-fluorobicyclo[1.1.1]pentan-1-yl}octan-1-ol and purified by columnchromatography on silica gel (PE/EtOAc 2:1) to give9-(oxetan-3-yl)nonan-1-ol (500 mg, 76%) as an oil.

Step 4: Synthesis of 9-(oxetan-3-yl)nonanal

Prepared in a manner similar to8-(3-fluorobicyclo[1.1.1]pentan-1-yl)octanal to give9-(oxetan-3-yl)nonanal (300 mg, 61%) as an oil, that was used directlyin the next step, without further purification.

Step 5: Synthesis of 25-(oxetan-3-yl)pentacos-16-enoic Acid

Prepared in a manner similar to2443-fluorobicyclo[1.1.1]pentan-1-yl)tetracos-16-enoic acid and purifiedby column chromatography on silica gel (PE/EtOAc 1:1) to give25-(oxetan-3-yl)pentacos-16-enoic acid (230 mg, 35%) as a solid. LC/MS:mass calcd. for C₂₈H₅₂O₃: 436, found: 435 [M+H]⁺.

Step 6: Synthesis of 25-(oxetan-3-yl)pentacosanoic Acid

Prepared in a manner similar to23-(3-fluorobicyclo[1.1.1]pentan-1-yl)tricosanoic acid give25-(oxetan-3-yl)pentacosanoic acid (230 mg, 100%) as a solid. LC/MS:mass calcd. for C₂₈H₅₄O₃: 438, found: 437 [M+H]⁺.

Step 7: Synthesis ofN-[(2S,3S,4R)-3,4-dihydroxy-1-{[(2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy}octadecan-2-yl]-25-(oxetan-3-yl)pentacosanamide

Prepared in a manner similar toN-[(2S,3S,4R)-3,4-dihydroxy-1-{([(2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy}octadecan-2-yl]-11-(3-methyloxetan-3-yl)undecanamide,except DMF/THF was used as solvent. Purified by column chromatography onsilica gel (DCM/MeOH 9:1) and washed with CH₃CN to giveN-[(2S,3S,4R)-3,4-dihydroxy-1-{[(2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy}octadecan-2-yl]-25-(oxetan-3-yl)pentacosanamide(28.4 mg, 13%). LC/MS: mass calcd. for C₅₂H₁₀₁NO₁₀: 899.74, found:922.80 [M+Na]⁺; ¹H NMR (400 MHz, CD₃OD) δ 4.79 (dd, J=7.8, 5.8 Hz, 2H),4.36 (t, J=6.1 Hz, 2H), 4.08-4.24 (m, 11H), 3.54-3.87 (m, 10H), 2.22 (t,J=7.4 Hz, 2H), 1.53-1.72 (m, 5H), 1.27-1.37 (m, 68H), 0.88-0.92 (m, 3H).

Synthesis ofN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-7-(dimethyl(octadecyl)silyl)heptanamide

Step 1: Synthesis of(7-(benzyloxy)hept-1-yn-1-yl)dimethyl(octadecyl)silane

To a mixture of ((hept-6-yn-1-yloxy)methyl)benzene (1.0 g, 4.9 mmol) inTHF (20 mL) at −78° C. under an atmosphere of N₂ was added n-BuLi, 2.5M, in hexane (2.2 mL, 5.4 mmol). The mixture was stirred at −78° C. for0.5 h, then chlorodimethyl(octadecyl)silane (1.7 g, 4.9 mmol) was addedand the mixture was warmed to rt and stirred overnight. H₂O (30 ml) wasadded and the mixture was extracted with EtOAc (50 mL×3). The combinedorganic layers were washed with brine (30 mL), dried over anhydrousNa₂SO₄, filtered and the filtrate was concentrated under reducedpressure. The residue was purified by column chromatography on silicagel (PE/EtOAc 3:1) to give(7-(benzyloxy)hept-1-yn-1-yl)dimethyl(octadecyl)silane (1.0 g, 39%) asan oil.

Step 2: Synthesis of 7-(dimethyl(octadecyl)silyl)heptan-1-ol

A mixture of [7-benzyloxy)hept-1-yn-1-yl]dimethyloctadecylsilane (1.0 g,1.95 mmol), Pd/C (100 mg), MeOH (6 mL) and EtOAc (18 mL) was stirred atrt under an atmosphere of H₂ (1 atm) for 16 h. The mixture was filteredthrough a pad of Celite and the filter cake was washed with MeOH. Thefiltrate was concentrated under reduced pressure to give7-(dimethyl(octadecyl)silyl)heptan-1-ol (300 mg, 36%) as a solid.

Step 3: Synthesis of 7-[dimethyl(octadecyl)silyl]heptanoic Acid

A mixture of CrO₃ (281 mg, 2.8 mmol), H2SO₄ (1 mL) and H₂O (2 mL) underan atmosphere of N₂ was stirred at rt for 10 min, then7-[dimethyl(octadecyl)silyl]heptan-1-ol (300 mg, 0.7 mmol), acetone (24mL) and DCM (8 mL) were added, and the mixture was stirred at rtovernight. The solvent was removed under reduced pressure, H₂O (30 mL)was added and the mixture was extracted with DCM (50 mL×4). The combinedorganic layers were washed with brine (30×3 mL), dried over Na₂SO₄,filtered and the filtrate was concentrated under reduced pressure. Theresidue was purified by column chromatography on silica gel (PE/EtOAc1:1) to give 7-[dimethyl(octadecyl)silyl]heptanoic acid (160 mg, 52%) asa solid.

Step 4: Synthesis ofN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-7-(dimethyl(octadecyl)silyl)heptanamide

Prepared in a manner similar toN-[(2S,3S,4R)-3,4-dihydroxy-1-{[(2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy}octadecan-2-yl]-11-(3-methyloxetan-3-yl)undecanamide.Purified by column chromatography on silica gel (DCM/MeOH 9:1) andwashed with CH₃CN to giveN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-7-(dimethyl(octadecyl)silyl)heptanamide(9 mg, 9%). LC/MS: mass calcd. for C₅₁H₁₀₃NO₉Si: 901.74, found: 924.75[M+Na]⁺, ¹H NMR (300 MHz, CD₃OD) δ 4.20 (d, J=6.2 Hz, 1H), 3.56-3.93 (m,10H), 2.24 (t, J=7.5 Hz, 2H), 1.53-1.70 (m, 5H), 1.19-1.41 (m, 61H),0.90-0.94 (m, 6H), 0.49-0.55 (m, 4H), 0.01 (s, 6H).

Synthesis ofN-[(2S,3S,4R)-3,4-dihydroxy-1-{[(2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy}octadecan-2-yl]-8-(heptadecyldimethylsilyl)octanamide

Step 1: Synthesis of[8-(benzyloxy)oct-1-yn-1-yl](heptadecyl)dimethylsilane

Prepared in a similar manner to(7-(benzyloxy)hept-1-yn-1-yl)dimethyl(octadecyl)silane and purified bycolumn chromatography on silica gel (PE/EtOAc 3:1) to give[8-(benzyloxy)oct-1-yn-1-yl](heptadecyl)dimethylsilane (800 mg, 38%) asan oil.

Step 2: Synthesis of 8-(heptadecyldimethylsilyl)octan-1-ol

Prepared in a manner similar to 7-(dimethyl(octadecyl)silyl)heptan-1-olto give 8-(heptadecyldimethylsilyl)octan-1-ol (150 mg, 36%) as a solid.

Step 3: Synthesis of 8-(heptadecyldimethylsilyl)octanoic Acid

Prepare in a manner similar to 7-[dimethyl(octadecyl)silyl]heptanoicacid. Purified by column chromatography on silica gel (PE/EtOAc 1:1) togive 8-(heptadecyldimethylsilyl)octanoic acid (80 mg, 52%) as a solid.

Step 4: Synthesis ofN-[(2S,3S,4R)-3,4-dihydroxy-1-{[(2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy}octadecan-2-yl]-8-(heptadecyldimethylsilyl)octanamide

Prepared in a manner similar toN-[(2S,3S,4R)-3,4-dihydroxy-1-{[(2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy}octadecan-2-yl]-11-(3-methyloxetan-3-yl)undecanamide.Purified by column chromatography on silica gel (DCM/MeOH 9:1) andwashed with CH₃CN to giveN-[(2S,3S,4R)-3,4-dihydroxy-1-{[(2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy}octadecan-2-yl]-8-(heptadecyldimethylsilyl)octanamide(11.9 mg, 9.0%). LC/MS: mass calcd. for C₅₁H₁₀₃NO₉Si: 901.74, found:924.85 [M+Na]⁺; ¹H NMR (400 MHz, CD₃OD) δ 4.19 (dt, J=6.9, 4.4 Hz, 1H),3.53-3.93 (m, 10H), 2.24 (t, J=7.5 Hz, 2H), 1.56-1.68 (m, 5H), 1.28-1.44(m, 61H), 0.90-0.94 (m, 6H), 0.51-0.55 (m, 4H), 0.01 (s, 6H).

Synthesis ofN-((1S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-24-(trimethylsilyl)tetracosanamide

Step 1: Synthesis of 8-(trimethylsilyl)octanal

To a mixture of 8-(trimethylsilyl)octan-1-ol [CAS No. 473844-91-6] (600mg, 2.96 mmol) and pyridine (467 mg, 5.93 mmol) in DCM (20 mL) was addedDess-Martin periodinane (1.88 g, 4.44 mmol). The mixture was stirred atrt for 3 h, then purified directly by column chromatography on silicagel (PE/EtOAc 5:1) to give 8-(trimethylsilyl)octanal (450 mg, 76%) as anoil.

Step 2: Synthesis of 24-(trimethylsilyl)tetracos-16-enoic Acid

To a mixture of (15-carboxypentadecyl)triphenylphosphonium bromide (995mg, 2.24 mmol), THF (10 mL) at 0° C. under an atmosphere of N₂ was added2 M NaHMDS (2.48 mL, 4.94 mmol). The mixture was warmed to rt andstirred for 1 h, then 8-(trimethylsilyl)octanal (450 mg, 2.24 mmol) wasadded and the mixture was stirred overnight at rt, then acidified topH˜6 with 1N HCl. The mixture was extracted with EtOAc (3×20 mL), thecombined organic layers were washed with brine (50 mL), dried overanhydrous Na₂SO₄, filtered and the filtrate was concentrated underreduced pressure. The residue was purified by column chromatography onsilica gel (PE/EtOAc 1:1) to give 24-(trimethylsilyl)tetracos-16-enoicacid (100 mg, 10%) as a solid. ¹H NMR (400 MHz, CDCl₃) δ 5.33-5.43 (m,2H), 2.37 (t, J=7.5 Hz, 2H), 2.04 (q, J=6.4 Hz, 4H), 1.65 (q, J=7.3 Hz,2H), 1.29 (d, J=10.0 Hz, 32H), 1.24 (s, 2H), 0.53-0.45 (m, 2H), 0.09 (s,9H).

Step 3: Synthesis of 24-(trimethylsilyl)tetracosanoic Acid

A mixture of 24-(trimethylsilyl)tetracos-16-enoic acid (100 mg, 0.22mmol). PtO₂ (2.5 mg) and EtOH (30 mL) were stirred under an atmosphereof H₂ (balloon) for 1 h, then filtered through a pad of Celite and thefiltrate was concentrated under reduced pressure to give24-(trimethylsilyl)tetracosanoic acid (100 mg, 100%) as a solid. ¹H NMR(400 MHz, CDCl₃) δ 2.23-2.36 (m, 2H), 1.55-1.60 (m, 2H), 1.27-1.32 (m,40H), 0.48-0.51 (m, 2H), 0.01 (s, 9H).

Step 4: Synthesis ofN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-24-(trimethylsilyl)tetracosanamide

Prepared in a manner similar toN-[(2S,3S,4R)-3,4-dihydroxy-1-{[(2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy}octadecan-2-yl]-11-(3-methyloxetan-3-yl)undecanamide,except DMF/THF was used as solvent. Purified by column chromatography onsilica gel (DCM/MeOH) and washed with CH₃CN to giveN-((2S,3S,4R)-3,4-dihydroxy-1-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-24-(trimethylsilyl)tetracosanamide(4.1 mg, 2.7%). LC/MS: mass calcd. for C₅₁H₁₀₃NO₉Si: 901.74, found:924.80 [M+Na]⁺; ¹H NMR (300 MHz, CD₃OD) δ 4.17 (d, J=6.2 Hz, 1H),3.54-3.93 (m, 10H), 2.22 (t, J=7.4 Hz, 2H), 1.50-1.69 (m, 6H), 1.26-1.39(m, 62H), 0.88 (t, J=8.8, 7.2 Hz, 3H), 0.45-0.53 (m, 2H), 0.01 (s, 9H).

Synthesis of1-allyl-4-((2S,3S,4R)-3,4-bis(benzyloxy)-1-(((2S,3R,4S,5S,6R),3,4,5-tris(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-1,4-dihydro-5H-tetrazol-5-one

Step 1: Synthesis of(2R,3S,4S,5R,6S)-3,4,5-tris(benzyloxy)-2-((benzyloxy)methyl)-6-(((2S,3S,4R)-3,4-bis(benzyloxy)-2-isocyanatooctadecyl)oxy)tetrahydro-2H-pyran

To a mixture of(2S,3S,4R)-3,4-bis(benzyloxy)-1-(((2S,3R,4S,5S,6R)-3,4,5-tris(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-amine(700 mg, 0.68 mmol) and Et₃N (84 mg, 0.83 mmol) in DCM (20 mL) at 0° C.was added triphosgene (224 mg, 0.75 mmol). The mixture was stirred at 0°C. for 1 h, then diluted with H₂O (20 mL) and extracted with DCM (3×30mL). The combined organic layers were dried over anhydrous Na₂SO₄,filtered and the filtrate was concentrated under reduced pressure togive(2R,3S,4S,5R,6S)-3,4,5-tris(benzyloxy)-2-((benzyloxy)methyl)-6-(((2S,3S,4R)-3,4-bis(benzyloxy)-2-isocyanatooctadecyl)oxy)tetrahydro-2H-pyran(700 mg, 98%) as a solid. LC/MS: mass calcd. for C₆₇H₈₃NO₉: 1045.61,found: 1068.50 [M+Na]⁺.

Step 2: Synthesis of1-((2S,3S,4R)-3,4-bis(benzyloxy)-1-(((2S,3R,4S,5S,6R)-3,4,5-tris(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-1,4-dihydro-5H-tetrazol-5-one

A mixture of(2R,3S,4S,5R,6S)-3,4,5-tris(benzyloxy)-2-((benzyloxy)methyl)-6-(((2S,3S,4R)-3,4-bis(benzyloxy)-2-isocyanatooctadecyl)oxy)tetrahydro-2H-pyran(700 mg, 0.67 mmol) and TMSN₃ (20 mL) was heated to 100° C., and stirredovernight. After cooling, the mixture was concentrated under the reducedpressure and the residue was purified by column chromatography on silicagel (PE/EtOAc 2:1) to give1-((2S,3S,4R)-3,4-bis(benzyloxy)-1-(((2S,3R,4S,5S,6R)-3,4,5-tris(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-1,4-dihydro-5H-tetrazol-5-one(400 mg, 55%) as a solid. LC/MS: mass calcd. for C₆₇H₈₄N₄O₉: 1088.62,found: 1111.50 [M+Na]⁺.

Step 3: Synthesis of1-allyl-4-((2S,3S,4R)-3,4-bis(benzyloxy)-1-(((2S,3R,4S,5S,6R)-3,4,5-tris(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-1,4-dihydro-5H-tetrazol-S-one

To a mixture of1-((2S,3S,4R)-3,4-bis(benzyloxy)-1-(((2S,3R,4S,5S,6R)-3,4,5-tris(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-1,4-dihydro-5H-tetrazol-5-one(50 mg, 0.05 mmol), K₂CO₃ (26 mg, 0.19 mmol) and DMA (3 mL) was added3-iodoprop-1-ene (12 mg, 0.071 mmol) in DMA (0.1 mL). The mixture washeated to 60° C., and stirred for 4 h, then diluted with EtOAc (60 mL)and the mixture washed with H₂O (4×15 mL), brine (2×15 mL), dried overNa₂SO₄, filtered and the filtrate was concentrated under reducedpressure to give1-allyl-4-((2S,3S,4R)-3,4-bis(benzyloxy)-1-(((2S,3R,4S,5S,6R)-3,4,5-tris(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-2-yl)oxy)octadecan-2-yl)-1,4-dihydro-5H-tetrazol-5-one(30 mg, 58%) as a solid. LC/MS: mass calcd. for C₇₀H₈₈N₄O₉: 1128.66,found: 1151.55 [M+Na]⁺.

Example 2—In Vitro Activation of Human iTCR Through a Jurkat ReporterCell Line

In order to determine the human iTCR activation potential induced by thecompounds described herein, a jurkat cell line (JiNKT) was transfectedwith the human iTCR, and GFP under the NFkB promoter (cell line licensedfrom the Medical University of Vienna). A BWS147 cell line (BWSTIM) wasalso transfected with CD80 and CD1d to act as the antigen-presentingcell.

Methods

DCD molecules or α-GalCer were both dissolved in DMSO at a 5 mg/mL stocksolution. BWSTIM cells were loaded with DCD molecules or α-GalCer atvarying concentrations for 4 hours at 37° C. at a concentration of 20kcells/well in 200 μL of media in a u-bottom 96 well dish. BWSTIM cellswere washed 2× with media, then incubated with JiNKT cells at aconcentration of 80k cells/well in a u-bottom 96 well dish Cells wereco-cultured for 18 to 24 hours. The percentage of cells expressing highlevels of GFP was measured through flow cytometry (after gating out themCD45+BWSTIM cells).

Results

FIG. 1A depicts results from the activation of human iTCR study with theJurkat reporter cell line. FIG. 1A depict the percentage GFP+ of cellsat different concentrations of compounds DCD-101, DCD-102, DCD-103,DCD-104, DCD-105, DCD-106, DCD-108, DCD-112, DCD-113, DCD-114, DCD115,and DCD-116, DCD118, DCD-119, DCD-120. DCD-121, DCD-122, DCD-123,DCD-124, DCD-125, DCD-126, DCD-127, DCD-128. DCD-129, DCD-130, DCD-131,DCD-132, DCD-133, DCD-134, DCD-135, DCD-136, DCD-137, DCD-138, DCD-139,DCD-140, DCD-141, DCD-142, DCD-143, DCD-144. DCD-145, DCD-146, DCD-147,DCD-148, DCD-149, DCD-150, DCD-151, DCD-152, DCD-153, DCD-154, DCD-155,DCD-156, DCD-157, DCD-158, and DCD-159. Alpha-galactosylceramide(α-GalCer) was also tested for comparison.

FIG. 1B shows DCD-127, DCD-141, DCD-143, DCD-144, DCD-136, DCD-133,DCD-122, DCD-155, DCD-118, DCD-121, DCD-101, DCD-138, DCD-153, DCD-119,DCD-149, DCD-150, DCD-139, DCD-125, DCD-148, DCD-103, DCD-113, DCD-106,DCD-114, DCD-151, DCD-137, DCD-128, DCD156, DCD-104, DCD-130, DCD-140,DCD-157 all had lower EC50s compared to α-GalCer

Conclusions

Compound DCD-127 demonstrated the highest % GFP+ at the lowestconcentration.

Example 3—In Vitro Activation of Mouse iTCR Through a DN3-a4.1.2 iNKTHybridoma Cell Line

In order to determine the mouse iTCR activation potential induced by thecompounds described herein, a mouse iNKT hybridoma cell line (DN3.2)from the La Jolla Institute for Allergy and Immunology were used as theiNKT. The BWS147 cell line (BWSTIM) with CD80 and CD1d acted as theantigen-presenting cell.

Methods

DCD molecules or α-GalCer were both dissolved in DMSO at a 5 mg/mL stocksolution. BWSTIM cells were loaded with DCD molecules or α-GalCer ofvarying concentrations for 4 hours at 37° C. at a concentration of 20kcells/well in 200 μL of media in a u-bottom 96 well dish. BWSTIM cellswere washed 2× with media, then incubated with DN3.2 cells at aconcentration of 80k cells/well in a u-bottom 96 well dish. Cells wereco-cultured for 48 hours. Media was collected and IL-2 was measuredusing the CisBio HTRF ELISA detection kit.

Results

FIG. 2A depicts results from the activation of the mouse iTCR activationstudy with the DN3.2 reporter cell line. The amount of IL-2 secretion inresponse to incubation with DCD-101, DCD-102, DCD-104, DCD105, DCD-106and α-GalCer is shown. No drug loading was also tested and is shown inFIG. 2A as well. FIG. 2A shows that when drug was loaded at aconcentration of 0.01 μg/mL, each of DCD-101, DCD-102 and DCD-106exhibited greater IL-2 secretion than α-GalCer.

Conclusions

Compounds DCD101, showed the highest IL-2 secretion at the lowestconcentrations.

Example 4—Cytokine Secretion of Primary Human iNKT Cells

In order to determine the activation profile induced by the compoundsdescribed herein, primary human iNKT cells were co-cultured withdrug-loaded BWSTIM cells

Methods

Day 0: PBMCs were isolated from human blood using the STEMCELLTECHNOLOGIES™ SepMate™ PBMC isolation system. iNKT cells were thenselected using the Miltenyi NKT magnetic cell separation kit.

Day 2: DCD molecules or α-GalCer were both dissolved in DMSO at a 5mg/mL stock solution. BWSTIM cells were fixed with mitomycin C, thenloaded with 10 μg/mL of DCD molecules or α-GalCer for 4 hours at 37° C.at a concentration of 20k cells/well in 200 μL of media in a u-bottom 96well dish. Cells were co-cultured with 80k 6B11+ selected primary humaniNKT cells.

D4: Media was collected. Cytokines were measured using the Satorious 4Plex kit on the iQue3 cytometer.

Results

FIG. 3A depicts the secretion of the cytokine interferon gamma (IFNγ) inresponse to activation by compounds DCD-101, DCD-104, DCD-106 andα-GalCer. FIG. 3B depicts the secretion of the cytokine interleukin-6(IL-6) in response to activation by compounds DCD-101, DCD-104, DCD-106and α-GalCer. FIG. 3C depicts the secretion of the cytokine tumornecrosis factor alpha (TNFα) in response to activation by compoundsDCD-101, DCD-104, DCD-106 and α-GalCer.

Conclusions

DCD-101, DCD-104, and DCD-106 all induce significantly higher levels ofIFNγ. IL6, and TNFα compared to the no-compound control. (**p<0.01;*p<0.05)

Example 5—In Vivo IFNγ Activation and iNKT Cell Expansion in C57BL/6JMice

In order to determine the expansion induced by the molecules describedherein, molecules were injected into C57BL/6J mice. Serum IFNγ levelsand the expansion of iNKT cells within splenocytes was measured 4 dayspost-IP injection.

Methods

Eight weeks old C57BL/6J mice were injected (I.P.) with 2 μg of α-GalCeror DCD molecules. Molecules were either dissolved in DMSO at a 5 mg/mLstock solution, or formulated into liposomes through thin-filmrehydration, then extrusion through 200 nm filters. Liposome-basedformulations were constructed using soy phosphatidylcholine,cholesterol, and DCD or α-GalCer in a 2.1:0.15 ratio. Twenty hourspost-injection, blood was collected from the tail to measure the levelof serum IFNγ using an ELISA kit from Biolegend. Four dayspost-injection, the mice were sacrificed and the splenocytes wereisolated. The percentage of iNKT cells within the spleens was measuredusing flow cytometry, selecting for live cells and mouse CD1d-α-GalCertetramer+ cells.

Results

FIG. 4A depicts the activation of immune cells, as measured by theamount of serum IFNγ using ELISA, in response to injection of thecompounds DCD-101, DCD-119, DCD-123, DCD125, DCD127, DCD-128, DCD-134,DCD-142, DCD-145, DCD-146, DCD-147, DCD-148, DCD-149, DCD-150, DCD-151,DCD-152, DCD-153, DCD-154, DCD-155, DCD-156, DCD-157, DCD-158, andDCD-159 with comparison to α-GalCer twenty hours after injection.

FIG. 4B depicts the expansion of iNKT cells in the mouse spleen inresponse to injection of the compounds DCD-101, DCD-104, DCD-106,DCD-119, DCD-142, DCD-145, DCD-146, DCD-147, DCD-148, DCD-149, DCD-150,DCD-151, DCD-152, DCD-153, DCD-154, DCD-155, DCD-156, DCD-157, DCD-158,and DCD-159 with comparison to α-GalCer.

Conclusions

The average serum IFNγ collected in the DCD-152, DCD-153, DCD-154,DCD-155.

DCD-156, and DCD-157 is all higher than the average serum IFNγ ofC57BL/6J mice treated with α-GalCer.

The average iNKT cell isolated from the spleen of C57BL/6J mice treatedwith DCD-153 or DCD-154 are both higher than mice treated with α-GalCer.

Example 6—Diet Induced Obesity (HFD) Mouse Model Study

Senescence is a feature of pre-adipocytes in obese individuals. To studythe efficacy of the molecules described herein in decreasing senescencein fat, a diet induced obesity high fat diet (HFD) mouse model was used.22-week-old HFD mice were injected with α-GalCer as a control andcompared with the compounds of the present disclosure. The blood andspleen (or adipose tissue) were collected to measure iNKT activation andexpansion, respectively. Cells in the eWAT (adipose tissue) were alsocollected and measured for % senescent cells. HFD mice were compared tonon-HFD (normal diet) mice.

Methods

22-week-old C57BL/6J mice on a chow or high-fat diet (HFD) were injected(I.P) with 2 μg of α-GalCer, compound DCD-101 or compound DCD-154.Molecules were either dissolved in DMSO at a 5 mg/mL stock solution, orformulated into liposomes through thin-film rehydration, then extrusionthrough 200 nm filters. Liposome-based formulations were constructedusing soy phosphatidylcholine, cholesterol, and DCD or α-GalCer in a2:1.0.15 ratio. Two or twenty hours post-injection, blood was collectedfrom the tail to measure the level of IFNγ using ELISA. Four dayspost-injection, mice were sacrificed to collect eWAT or spleen. Spleenwas used to measure the number of iNKT cells, and eWAT was used tomeasure the number of iNKT cells and senescent cells using flowcytometry, iNKT cells were identified in the digested adipose tissue bygating live cells and mouse CD1d-α-GalCer tetramer+ cells. Senescentcells were measured within the processes adipose tissue by selectingmCD45− cells, and C12FDG^(HIGH) cells.

Results

FIG. 5A depicts the expansion of immune cells in the spleen of: 1) miceon a normal diet; 2) mice on a high fat diet injected with diluent; 3)mice on a high fat diet injected with α-GalCer and 4) mice on a high fatdiet injected with compound DCD-101. Both α-GalCer and compound DCD-101caused expansion of immune cells in HFD mouse spleen, at four dayspost-injection.

FIG. 5B depicts the expansion of immune cells in the eWAT of: 1) mice ona normal diet; 2) mice on a high fat diet injected with diluent; 3) miceon a high fat diet injected with α-GalCer and 4) mice on a high fat dietinjected with compound DCD-154 Both α-GalCer and compound DCD-154 causedexpansion of immune cells in HFD mouse eWAT, as measured by flowcytometry of the percent iNKT cells of live cells in the mouse eWAT.

FIG. 5C depicts the activation of immune cells, as measured by thelevels of serum IFNγ using ELISA. Both α-GalCer and DCD-101significantly increased levels of IFNγ in the HFD model two hours postinjection.

FIG. 5D depicts the activation of immune cells, as measured by thesecretion of IFNγ using ELISA. Both α-GalCer and DCD-154 significantlyincreased levels of IFNγ in the HFD model twenty hours post injection.

Non-immune C12FDG+ cells from eWAT were identified via flow cytometry.Decrease in the number of C12FDG positive cells indicate a decrease inthe number of senescent cells in eWAT. HFD mice treated with DCD-101,DCD-154 and α-GalCer were effective in decreasing the accumulation ofsenescent cells in eWAT. (FIG. 5E). (****p<0.0001; ***P<0.001, **p<0.01)

Conclusions

In the HFD mouse model, DCD-101 and DCD-154 expand iNKT cells in thespleen or adipose tissue respectively, 4 days post-treatment. BothDCD-101 and DCD-154 induce secretion of IFNγ between two and twentyhours post-treatment. Senescent cell reduction in adipose tissue isobserved in both DCD-101 and DCD-154.

Example 7—Inactivity of Compounds in In Vitro Studies with Human andMouse iTCR in Jurkat Reporter and DN3-a4.1.2 iNKT Hybridoma Cell Lines

A jurkat cell line (JiNKT) was transfected with the human iTCR, and GFPunder the NFkB promoter. A BWS147 cell line (BWSTIM) was alsotransfected with CD80 and CD1d to act as the antigen-presenting cell. Amouse iNKT hybridoma cell line (DN3.2) was also used as the iNKT. TheBWS147 cell line (BWSTIM) with CD80 and CD1d acted as theantigen-presenting cell.

Compounds Tested:

GFP Expression Studies

Methods

GVK molecules or α-GalCer were both dissolved in DMSO at a 5 mg/mL stocksolution. BWSTIM cells were loaded with compounds for 4 hours at 37° C.at a concentration of 20k cells/well in 200 μL of media in a u-bottom 96well dish. BWSTIM cells were washed 2× with media, then incubated withJiNKT cells at a concentration of 80k cells/well in a u-bottom 96 welldish. Cells were co-cultured for 24 hours. The percentage of cellsexpressing high levels of GFP was measured through flow cytometry. 1μg/mL of molecule GVK1a, GVK1b, GVK1c, or GVK1f was incubated withBWSTIM+JiNKT, and compared to α-GalCer.

Results

As shown in FIG. 6A, compound GVK1a, GVK1b, GVK1c, and GVK1f did notinduce GFP expression higher than the negative control, while α-GalCerinduction remained high. (****p<0.000)

IL-2 Expression Studies

Methods

GVK molecules or α-GalCer were both dissolved in DMSO at a 5 mg/mL stocksolution. BWSTIM cells were loaded with compounds for 4 hours at 37° C.at a concentration of 20k cells/well in 200 μL of media in a u-bottom 96well dish. BWSTIM cells were washed 2× with media, then incubated withDN3.2 cells at a concentration of 80k cells/well in a u-bottom 96 welldish. Cells were co-cultured for 48 hours. Media was collected and IL-2was measured using the CisBio HTRF ELISA detection kit.

Results

FIG. 6B depicts IL-2 expression by compounds GVK1a, GVK1b and GVK1f inthe DN3.2 cell line when loaded on BWSTIM CD₁d. As shown in FIG. 6B,each of compounds GVK1b, GVK1d and GVK1f did not induce expression ofIL-2. (****p<0.0001)

Cytokine Secretion Studies

In order to determine the activation profile induced by the compoundsdescribed herein, primary human iNKT cells were co-cultured withdrug-loaded BWSTIM cells.

Methods

GVK molecules or α-GalCer were both dissolved in DMSO at a 5 mg/mL stocksolution. BWSTIM cells were loaded with compounds for 4 hours at 37° C.at a concentration of 20k cells/well in 200 μL of media in a u-bottom 96well dish. Cells were co-cultured with 100k 6B11+ selected primary humaniNKT cells. Media was collected 2 days later. Cytokines were measuredusing the Satorious 4 Plex kit on the iQue3 cytometer.

Results

FIG. 6C depicts the secretion of the cytokine interferon gamma (IFNγ),tumor necrosis factor alpha (TNFα), interleukin-4 (IL-4) andinterleukin-6 (IL-6) in response to incubation with compounds GVK1a,GVK1b, GVK1c and GVK1f and α-GalCer. The secretion was compared tosecretion by cells in the absence of drug loading as a negative control.As shown in FIG. 8F, loading with each of compounds GVK1a, GVK1b, GVK1cand GVK1f did not increase secretion of IFNγ, TNF, IL-4 or IL-6 ascompared to cells in the absence of drug loading. Loading with α-GalCerexhibited a significantly greater increase in the secretion of each ofIFNγ, TNFα, IL4 or IL-6 as compared with GVK1a GVK1b, GVK1c and GVK1f.(****p<0.0001; *p<0.05)

TABLE 1 Summary of Experiments with Compounds GVK1a, GVK1b, GVK1c andGVK1f % GFP hiNKT % GFP hiNKT binding binding (recombinant (BWSTIM +protein IL-2 Secretion Primary iNKT JiNKT) CD1d + (A20 + cytokineCompound 1 μg/mL JiNKT) DN3.2) release α-GalCer %40-70 %30 GVK1a %<5away from %<5 away from Activated No activation of negative controlnegative control DN3.2 primary iNKT to release IL-2 to release IL-4,IL-6, IFNγ or TNFα cytokines GVK1b %<5 away from %<5 away from Does notNo activation of negative control negative control activate DN3.2primary iNKT to release IL-2 to release IL-4, IL-6, IFNγ or TNFαcytokines GVK1c %<5 away from %<5 away from Does not No activation ofnegative control negative control activate DN3.2 primary iNKT to releaseIL-2 to release IL-4, IL-6, IFNγ or TNFα cytokines GVK1f %<5 away from%<5 away from Does not No activation of negative control negativecontrol activate DN3.2 primary iNKT to release IL-2 to release IL-4,IL-6, IFNγ or TNFα cytokines

Example 8—Activation of iNKT Cells Selectively Kills Senescent Cells InVitro

Selective reduction in the presence of senescent cells by iNKT mediatedkilling in an in vitro sample was demonstrated using α-GalCer. HumaniNKT cells were isolated and activated by incubation with α-GalCer.Activated iNKT cells were combined with samples containing healthy cellsand senescent cells. As shown in FIGS. 7A, incubation of senescent cellswith activated iNKT cells resulted in a reduction in the presence ofsenescent cells over time whereas non-senescent cells were maintained.FIG. 7B demonstrates that senescent cells were selectively killedwhereas non-senescent cells were maintained in the presence of activatediNKT cells when evaluated after 8 hours and 18 hours.

Although the foregoing invention has been described in some detail byway of illustration and example for purposes of clarity ofunderstanding, it is readily apparent to those of ordinary skill in theart in light of the teachings of this invention that certain changes andmodifications may be made thereto without departing from the spirit orscope of the appended claims.

Accordingly, the preceding merely illustrates the principles of theinvention. It will be appreciated that those skilled in the art will beable to devise various arrangements which, although not explicitlydescribed or shown herein, embody the principles of the invention andare included within its spirit and scope. Furthermore, all examples andconditional language recited herein are principally intended to aid thereader in understanding the principles of the invention and the conceptscontributed by the inventors to furthering the art, and are to beconstrued as being without limitation to such specifically recitedexamples and conditions. Moreover, all statements herein recitingprinciples, aspects, and embodiments of the invention as well asspecific examples thereof, are intended to encompass both structural andfunctional equivalents thereof. Additionally, it is intended that suchequivalents include both currently known equivalents and equivalentsdeveloped in the future. i.e., any elements developed that perform thesame function, regardless of structure. Moreover, nothing disclosedherein is intended to be dedicated to the public regardless of whethersuch disclosure is explicitly recited in the claims.

The scope of the present invention, therefore, is not intended to belimited to the exemplary embodiments shown and described herein. Rather,the scope and spirit of present invention is embodied by the appendedclaims. In the claims, 35 U.S.C. § 112(f) or 35 U.S.C. § 112(6) isexpressly defined as being invoked for a feature in the claim only whenthe exact phrase “means for” or the exact phrase “step for” is recitedat the beginning of such feature in the claim; if such exact phrase isnot used in a feature in the claim, then 35 U.S.C. § 112(f) or 35 U.S.C.§ 112(6) is not invoked.

1-15. (canceled)
 16. A compound selected from the group consisting of:

or a pharmaceutically acceptable salt thereof.
 17. The compound of claim16, or a pharmaceutically acceptable salt thereof, wherein the compoundis:

or a pharmaceutically acceptable salt thereof.
 18. The compound of claim16, or a pharmaceutically acceptable salt thereof, wherein the compoundis:

or a pharmaceutically acceptable salt thereof.
 19. The compound of claim16, or a pharmaceutically acceptable salt thereof, wherein the compoundis:

or a pharmaceutically acceptable salt thereof.
 20. The compound of claim16, or a pharmaceutically acceptable salt thereof, wherein the compoundis:

or a pharmaceutically acceptable salt thereof.
 21. The compound of claim16, or a pharmaceutically acceptable salt thereof, wherein the compoundis:

or a pharmaceutically acceptable salt thereof.
 22. The compound of claim16, or a pharmaceutically acceptable salt thereof, wherein the compoundis:

acceptable salt thereof.
 23. The compound of claim 16, or apharmaceutically acceptable salt thereof, wherein the compound is:

or a pharmaceutically acceptable salt thereof.
 24. The compound of claim16, or a pharmaceutically acceptable salt thereof, wherein the compoundis:

or a pharmaceutically acceptable salt thereof.
 25. A pharmaceuticalcomposition, comprising: a compound of claim 16, or a pharmaceuticallyacceptable salt thereof; and at least one pharmaceutically acceptablecarrier.
 26. A pharmaceutical composition, comprising: a compound ofclaim 17, or a pharmaceutically acceptable salt thereof, and at leastone pharmaceutically acceptable carrier.
 27. A pharmaceuticalcomposition, comprising: a compound of claim 18, or a pharmaceuticallyacceptable salt thereof; and at least one pharmaceutically acceptablecarrier.
 28. A pharmaceutical composition, comprising: a compound ofclaim 19, or a pharmaceutically acceptable salt thereof; and at leastone pharmaceutically acceptable carrier.
 29. A pharmaceuticalcomposition, comprising: a compound according to claim 20, or apharmaceutically acceptable salt thereof; and at least onepharmaceutically acceptable carrier.
 30. A pharmaceutical composition,comprising: a compound of claim 21, or a pharmaceutically acceptablesalt thereof; and at least one pharmaceutically acceptable carrier. 31.A pharmaceutical composition, comprising: a compound of claim 22, or apharmaceutically acceptable salt thereof; and at least onepharmaceutically acceptable carrier.
 32. A pharmaceutical composition,comprising: a compound of claim 23, or a pharmaceutically acceptablesalt thereof; and at least one pharmaceutically acceptable carrier. 33.A pharmaceutical composition, comprising: a compound of claim 24, or apharmaceutically acceptable salt thereof; and at least onepharmaceutically acceptable carrier.
 34. A method for selectivelyreducing the presence of or eliminating senescent cells by activatinginvariant natural killer T (iNKT) cells in a subject in need thereof,comprising: administering to the subject a compound of claim 16, or apharmaceutically acceptable salt thereof.
 35. A method of treating adisease, disorder or condition by activating invariant natural killer T(iNKT) cells in a subject in need thereof, comprising: administering tothe subject a compound of claim 16, or a pharmaceutically acceptablesalt thereof.
 36. The method of claim 35, wherein the disease, disorderor condition is an autoimmune disease, an allergic disease, a metabolicsyndrome or disorder, cancer, a pathogen infection, an eye disease, adisease of aging, fibrosis, heart disease, or kidney disease, or anycombination thereof.
 37. The method of claim 36, wherein the fibrosis islung fibrosis.
 38. The method of claim 36, wherein the fibrosis iskidney fibrosis.
 39. The method of claim 36, wherein the fibrosis isliver fibrosis.
 40. The method of claim 35, wherein the disease,disorder or condition is Type 1 diabetes, Type 2 diabetes, NAFLD, NASH,Rheumatoid Arthritis, Ulcerative Colitis, Multiple Sclerosis, FamilialHypercholesteremia, Giant Cell Arteritis, Idiopathic Pulmonary Fibrosis(IPF), idSystemic Lupus Erythematosus, Cachexia, Glaucoma, ChronicObstructive Pulmonary Disease, Systemic Sclerosis, Pulmonary ArterialHypertension, Lipodystrophy, Sarcopenia, Alopecia, Post MyocardialInfarction, Vitiligo, POTS, MCAD, Sjogren's, Scleroderma, HashimotoDisease, Ankylosing Spondylitis, Fibromyalgia, Sarcoidosis, Hepatitis,Raynauld's Syndrome, Mold Illness, Celiac, Crohn's, Pemphigus, SPS, PBC,Psoriatic Arthritis, CIDP, motor neuron disease, GPA, ALS, MyasenthiaGravis, or Presbyopia, or any combination thereof.
 41. The method ofclaim 35, wherein the disease, disorder or condition is Type 1 diabetes.42. The method of claim 35, wherein the disease, disorder or conditionis Type 2 diabetes.
 43. The method of claim 35, wherein the disease,disorder or condition is Idiopathic Pulmonary Fibrosis (IPF).